Organic compositions to treat hsf1-related diseases

ABSTRACT

The present disclosure relates to methods of treating heat shock factor 1 (HSF1)-related diseases such as cancer and viral diseases, using a therapeutically effective amount of a RNAi agent to HSF.

This application is a divisional application of U.S. Utility patentapplication Ser. No. 12/970,268 filed 16 Dec. 2010, which claimspriority to U.S. Provisional Application Ser. No. 61/228,137 filed 18Dec. 2009, the contents of which are incorporated herein by reference intheir entirety.

BACKGROUND OF THE INVENTION

HSF1 is the master regulator of the heat shock response, in whichmultiple genes are induced in response to temperature increase and otherstresses. At non-shock temperatures in humans and other vertebrates,HSF1 is produced constitutively, but is inactive and bound by proteinHSP90. At an elevated temperature, HSF1 is released by HSP90, moves fromthe cytoplasm to the nucleus, and trimerizes. This active HSF1 binds toheat shock elements (HSE) in DNA and activates transcription of heatshock genes by RNA polymerase II. The HSE has a consensus sequence ofthree repeats of NGAAN and is present in the promoter regions of theHSP90, HSP70 and HSP27 genes. During cessation of the heat shockresponse, HSF1 is phosphorylated by mitogen-activated protein kinases(MAPKs) and glycogen synthase kinase 3 (GSK3) and returns to an inactivestate. The biochemistry of HSF1 is described, inter alia, in Chu et al.1996 J. Biol. Chem. 271:30847-30857; Huang et al. 1997 J. Biol. Chem.272:26009-26016; and Morimoto et al. 1998 Nat. Biotech. 16: 833-838.

HSF1 interacts with additional factors. HSF1 binds to DNA-dependentprotein kinase (DNA-PK), which is involved in DNA repair. HSF1 is atarget of mitogen-activated protein kinases, and its activity isdown-regulated when the RAS signaling cascade is active.

Additional heat shock factor proteins in humans include HSF2, HSF3, andHSF4. HSF1, HSF2, and HSF3 are positive regulators of heat shock geneexpression, while HSF4 is a negative regulator. HSF1, HSF2 and HSF4 playa role in transcriptional control of other heat shock proteins. Thevarious HSF proteins share about 40% sequence identity.

HSF1 has been implicated in several diseases, including cancer and viraldiseases. HSF1 and other heat shock proteins (whose expression isincreased by HSF1) are over-expressed in, or have otherwise beenimplicated in breast, endometrial, fibrosarcoma, gastric, kidney, liver,lung, lymphoma, neuroectodermal, neuroblastoma, Ewing's sarcoma,prostate, skin, squamous cell, and testicular cancers, leukemia (e.g.,promyelocytic leukemia), and Hodgkin's disease.

Without wishing to be bound by any particular theory, it is believedthat heat shock proteins (HSP) may block the pathways of apoptosis andpermit malignant cells to arise despite the triggering of apoptoticsignals during transformation. HSP expression may also afford protectionto cancer cells from treatments such as chemotherapy and hyperthermia bythwarting the pro-apoptotic influence of these modalities.

Because HSF1 positively regulates HSPs, a need exists for therapeuticsthat modulate HSF1.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 illustrates various 5′-end modifications of RNAi agents to HSF1.

BRIEF SUMMARY OF THE INVENTION

The present disclosure encompasses RNAi agents to HSF1, useful intreatment of HSF1-related disease, such as cancer and viral diseases.

The present disclosure provides RNAi agents directed to the HSF1 (heatshock factor 1) gene. HSF1 is the master regulator of the heat shockresponse, in which multiple genes are induced in response to temperatureincrease and other stresses.

HSF1 has been implicated in several HSF1-related diseases, includingcancer and viral diseases. HSF1 and other heat shock proteins (whoseexpression is increased by HSF1) are over-expressed in, or haveotherwise been implicated in breast, endometrial, fibrosarcoma, gastric,kidney, liver, lung, lymphoma, neuroectodermal, neuroblastoma, Ewing'ssarcoma, prostate, skin, squamous cell, and testicular cancers, leukemia(e.g., promyelocytic leukemia), and Hodgkin's disease.

Because HSF1 positively regulates HSPs, a need exists for therapeuticsthat modulate HSF1. The RNAi agents of the present disclosure arespecific to HSF1 and can reduce expression of HSF1. These RNAi agentsare therefore useful in treating cancer and viral diseases.

DETAILED DESCRIPTION OF THE INVENTION

The present disclosure encompasses RNAi agents to HSF1, useful intreatment of HSF1-related disease, such as cancer and viral diseases.

Various Embodiments of the Disclosure Include the Following.

An RNAi Agent Comprising an Antisense Strand of an RNAi Agent DescribedHerein.

In one embodiment, the present disclosure relates to a compositioncomprising an

RNAi agent comprising an antisense strand, wherein the antisense strandcomprises at least 15 contiguous nucleotides differing by 0, 1, 2, or 3nucleotides from the antisense strand of an RNAi agent to HSF1 selectedfrom any sequence provided in a table herein (e.g., Table 1, Table 2,Table 3, Table 3A, Table 8, Table 9A, Table 9B, etc.). In anotherembodiment, the present disclosure relates to a composition comprisingan RNAi agent comprising a sense strand and an antisense strand, whereinthe antisense strand comprises at least 15 contiguous nucleotidesdiffering by 0, 1, 2, or 3 nucleotides from the antisense strand of anRNAi agent to HSF1 from any sequence provided herein. In anotherembodiment, the present disclosure relates to a composition comprisingan RNAi agent comprising a sense strand and an antisense strand, whereinthe sense strand comprises at least 15 contiguous nucleotides differingby 0, 1, 2, or 3 nucleotides from the sense strand and the antisensestrand comprises at least 15 contiguous nucleotides differing by 0, 1,2, or 3 nucleotides from the antisense strand of an RNAi agent to HSF1listed immediately above.

Particular duplexes include the following, wherein each duplex comprisesa set of SEQ ID NOs, wherein the first SEQ ID NO corresponds to thesense strand and the second SEQ ID NO corresponds to the antisensestrand: AD-20403 (SEQ ID NO: 131 and 643; or SEQ ID NO: 1155 and 1667);AD-20437 (SEQ ID NO: 166 and 678; or SEQ ID NO: 1190 and 1702); AD-20438(SEQ ID NO: 167 and 679; or SEQ ID NO: 1191 and 1703); AD-20439 (SEQ IDNO:

168 and 680; or SEQ ID NO: 1192 and 1704); AD-20487 (SEQ ID NO: 169 and681; or SEQ ID NO: 1193 and 1705); AD-20489 (SEQ ID NO: 171 and 683; orSEQ ID NO: 1195 and 1707); AD-20490 (SEQ ID NO: 172 and 684; or SEQ IDNO: 1196 and 1708); AD-20491 (SEQ ID NO: 173 and 685; or SEQ ID NO: 1197and 1709); AD-20548 (SEQ ID NO: 234 and 746; or SEQ ID NO: 1258 and1770); AD-20560 (SEQ ID NO: 269 and 781; or SEQ ID NO: 1293 and 1805);AD-20562 (SEQ ID NO: 271 and 783; or SEQ ID NO: 1295 and 1807); AD-20563(SEQ ID NO: 272 and 784; or SEQ ID NO: 1296 and 1808); AD-20564 (SEQ IDNO: 273 and 785; or SEQ ID NO: 1297 and 1809); AD-20578 (SEQ ID NO: 285and 797; or SEQ ID NO: 1309 and 1821); AD-20626 (SEQ ID NO: 290 and 802;or SEQ ID NO: 1314 and 1826); AD-20627 (SEQ ID NO: 291 and 803; or SEQID NO: 1315 and 1827); AD-20644 (SEQ ID NO: 308 and 820; or SEQ ID NO:1332 and 1844); AD-20648 (SEQ ID NO: 312 and 824; or SEQ ID NO: 1336 and1848); AD-20652 (SEQ ID NO: 316 and 828; or SEQ ID NO: 1340 and 1852);AD-20660 (SEQ ID NO: 324 and 836; or SEQ ID NO: 1348 and 1860); AD-20694(SEQ ID NO: 377 and 889; or SEQ ID NO: 1401 and 1913); AD-20707 (SEQ IDNO: 393 and 905; or SEQ ID NO: 1417 and 1929); AD-20730 (SEQ ID NO: 415and 927; or SEQ ID NO: 1439 and 1951); AD-20437.4 (SEQ ID NO: 3218 andSEQ ID NO: 3219); AD-20487.7 (SEQ ID NO: 3220 and SEQ ID NO: 3221);AD-20489.2 (SEQ ID NO: 3222 and SEQ ID NO: 3223); AD-20560.4 (SEQ ID NO:3224 and SEQ ID NO: 3225); AD-37718.1 (SEQ ID NO: 3226 and SEQ ID NO:3227); AD-37719.1 (SEQ ID NO: 3242 and SEQ ID NO: 3243); AD-37721.1 (SEQID NO: 3228 and SEQ ID NO: 3229); AD-37722.1 (SEQ ID NO: 3244 and SEQ IDNO: 3245); AD-37724.1 (SEQ ID NO: 3230 and SEQ ID NO: 3231); AD-37725.1(SEQ ID NO: 3246 and SEQ ID NO: 3247); AD-37727.1 (SEQ ID NO: 3232 andSEQ ID NO: 3233); AD-37728.1 (SEQ ID NO: 3248 and SEQ ID NO: 3249);AD-37730.1 (SEQ ID NO: 3234 and SEQ ID NO: 3235); AD-37733.1 (SEQ ID NO:3236 and SEQ ID NO: 3237); AD-37736.1 (SEQ ID NO: 3238 and SEQ ID NO:3239); AD-37740.1 (SEQ ID NO: 3240 and SEQ ID NO: 3241); AD-36969.2 (SEQID NO: 3250 and SEQ ID NO: 3251); AD-30071.2 (SEQ ID NO: 3252 and SEQ IDNO: 3253); AD-36970.2 (SEQ ID NO: 3254 and SEQ ID NO: 3255); AD-37739.1(SEQ ID NO: 3256 and SEQ ID NO: 3257); AD-37731.1 (SEQ ID NO: 3258 andSEQ ID NO: 3259); AD-37734.1 (SEQ ID NO: 3260 and SEQ ID NO: 3261);AD-37737.1 (SEQ ID NO: 3262 and SEQ ID NO: 3263); AD-37741.1 (SEQ ID NO:3264 and SEQ ID NO: 3265); AD-37720.1 (SEQ ID NO: 3266 and SEQ ID NO:3267); AD-37723.1 (SEQ ID NO: 3268 and SEQ ID NO: 3269); AD-37726.1 (SEQID NO: 3270 and SEQ ID NO: 3271); AD-37729.1 (SEQ ID NO: 3272 and SEQ IDNO: 3273); AD-37732.1 (SEQ ID NO: 3274 and SEQ ID NO: 3275); AD-37735.1(SEQ ID NO: 3276 and SEQ ID NO: 3277); AD-37738.1 (SEQ ID NO: 3278 andSEQ ID NO: 3279); AD-37742.1 (SEQ ID NO: 3280 and SEQ ID NO: 3281);AD-20303 (SEQ ID NO: 30 and 542; or SEQ ID NO: 1054 and 1566); AD-20313(SEQ ID NO: 40 and 552; or SEQ ID NO: 1064 and 1576); AD-20315 (SEQ IDNO: 42 and 554; or SEQ ID NO: 1066 and 1578); AD-20348 (SEQ ID NO: 56and 568; or SEQ ID NO: 1080 and 1592); AD-20362 (SEQ ID NO: 70 and 582;or SEQ ID NO: 1094 and 1606); AD-20364 (SEQ ID NO: 72 and 584; or SEQ IDNO: 1096 and 1608); AD-20365 (SEQ ID NO: 73 and 585; or SEQ ID NO: 1097and 1609); AD-20366 (SEQ ID NO: 74 and 586; or SEQ ID NO: 1098 and1610); AD-20373 (SEQ ID NO: 81 and 593; or SEQ ID NO: 1105 and 1617);AD-20376 (SEQ ID NO: 84 and 596; or SEQ ID NO: 1108 and 1620); AD-20378(SEQ ID NO: 85 and 597; or SEQ ID NO: 1109 and 1621); AD-20386 (SEQ IDNO: 93 and 605; or SEQ ID NO: 1117 and 1629); AD-20389 (SEQ ID NO: 117and 629; or SEQ ID NO: 1141 and 1653); AD-20391 (SEQ ID NO: 119 and 631;or SEQ ID NO: 1143 and 1655); AD-20392 (SEQ ID NO: 120 and 632; or SEQID NO: 1144 and 1656); AD-20397 (SEQ ID NO: 125 and 637; or SEQ ID NO:1149 and 1661); AD-20398 (SEQ ID NO: 126 and 638; or SEQ ID NO: 1150 and1662); AD-20399 (SEQ ID NO: 127 and 639; or SEQ ID NO: 1151 and 1663);AD-20401 (SEQ ID NO: 129 and 641; or SEQ ID NO: 1153 and 1665); AD-20402(SEQ ID NO: 130 and 642; or SEQ ID NO: 1154 and 1666); AD-20404 (SEQ IDNO: 132 and 644; or SEQ ID NO: 1156 and 1668); AD-20406 (SEQ ID NO: 136and 648; or SEQ ID NO: 1160 and 1672); AD-20407 (SEQ ID NO: 137 and 649;or SEQ ID NO: 1161 and 1673); AD-20408 (SEQ ID NO: 138 and 650; or SEQID NO: 1162 and 1674); AD-20409 (SEQ ID NO: 139 and 651; or SEQ ID NO:1163 and 1675); AD-20410 (SEQ ID NO: 140 and 652; or SEQ ID NO: 1164 and1676); AD-20411 (SEQ ID NO: 141 and 653; or SEQ ID NO: 1165 and 1677);AD-20413 (SEQ ID NO: 2042 and 2043; or SEQ ID NO: 2046 and 2047);AD-20422 (SEQ ID NO: 151 and 663; or SEQ ID NO: 1175 and 1687); AD-20428(SEQ ID NO: 157 and 669; or SEQ ID NO: 1181 and 1693); AD-20434 (SEQ IDNO: 163 and 675; or SEQ ID NO: 1187 and 1699); AD-20435 (SEQ ID NO: 164and 676; or SEQ ID NO: 1188 and 1700); AD-20488 (SEQ ID NO: 170 and 682;or SEQ ID NO: 1194 and 1706); AD-20493 (SEQ ID NO: 175 and 687; or SEQID NO: 1199 and 1711); AD-20495 (SEQ ID NO: 177 and 689; or SEQ ID NO:1201 and 1713); AD-20502 (SEQ ID NO: 184 and 696; or SEQ ID NO: 1208 and1720); AD-20507 (SEQ ID NO: 189 and 701; or SEQ ID NO: 1213 and 1725);AD-20513 (SEQ ID NO: 195 and 707; or SEQ ID NO: 1219 and 1731); AD-20527(SEQ ID NO: 209 and 721; or SEQ ID NO: 1233 and 1745); AD-20535 (SEQ IDNO: 217 and 729; or SEQ ID NO: 1241 and 1753); AD-20544 (SEQ ID NO: 230and 742; or SEQ ID NO: 1254 and 1766); AD-20545 (SEQ ID NO: 231 and 743;or SEQ ID NO: 1255 and 1767); AD-20546 (SEQ ID NO: 232 and 744; or SEQID NO: 1256 and 1768); AD-20547 (SEQ ID NO: 233 and 745; or SEQ ID NO:1257 and 1769); AD-20549 (SEQ ID NO: 235 and 747; or SEQ ID NO: 1259 and1771); AD-20552 (SEQ ID NO: 238 and 750; or SEQ ID NO: 1262 and 1774);AD-20555 (SEQ ID NO: 241 and 753; or SEQ ID NO: 1265 and 1777); AD-20556(SEQ ID NO: 242 and 754; or SEQ ID NO: 1266 and 1778); AD-20557 (SEQ IDNO: 243 and 755; or SEQ ID NO: 1267 and 1779); AD-20558 (SEQ ID NO: 267and 779; or SEQ ID NO: 1291 and 1803); AD-20561 (SEQ ID NO: 270 and 782;or SEQ ID NO: 1294 and 1806); AD-20565 (SEQ ID NO: 274 and 786; or SEQID NO: 1298 and 1810); AD-20566 (SEQ ID NO: 275 and 787; or SEQ ID NO:1299 and 1811); AD-20572 (SEQ ID NO: 280 and 792; or SEQ ID NO: 1304 and1816); AD-20574 (SEQ ID NO: 2044 and 2045; or SEQ ID NO: 2048 and 2049);AD-20575 (SEQ ID NO: 282 and 794; or SEQ ID NO: 1306 and 1818); AD-20577(SEQ ID NO: 284 and 796; or SEQ ID NO: 1308 and 1820); AD-20579 (SEQ IDNO: 286 and 798; or SEQ ID NO: 1310 and 1822); AD-20625 (SEQ ID NO: 289and 801; or SEQ ID NO: 1313 and 1825); AD-20633 (SEQ ID NO: 297 and 809;or SEQ ID NO: 1321 and 1833); AD-20634 (SEQ ID NO: 298 and 810; or SEQID NO: 1322 and 1834); AD-20640 (SEQ ID NO: 304 and 816; or SEQ ID NO:1328 and 1840); AD-20646 (SEQ ID NO: 310 and 822; or SEQ ID NO: 1334 and1846); AD-20650 (SEQ ID NO: 314 and 826; or SEQ ID NO: 1338 and 1850);AD-20653 (SEQ ID NO: 317 and 829; or SEQ ID NO: 1341 and 1853); AD-20661(SEQ ID NO: 325 and 837; or SEQ ID NO: 1349 and 1861); AD-20671 (SEQ IDNO: 337 and 849; or SEQ ID NO: 1361 and 1873); AD-20693 (SEQ ID NO: 376and 888; or SEQ ID NO: 1400 and 1912); AD-20700 (SEQ ID NO: 383 and 895;or SEQ ID NO: 1407 and 1919); AD-20702 (SEQ ID NO: 385 and 897; or SEQID NO: 1409 and 1921); AD-20709 (SEQ ID NO: 394 and 906; or SEQ ID NO:1418 and 1930); AD-20710 (SEQ ID NO: 395 and 907; or SEQ ID NO: 1419 and1931); AD-20714 (SEQ ID NO: 399 and 911; or SEQ ID NO: 1423 and 1935);AD-20716 (SEQ ID NO: 401 and 913; or SEQ ID NO: 1425 and 1937); AD-20728(SEQ ID NO: 413 and 925; or SEQ ID NO: 1437 and 1949); AD-20741 (SEQ IDNO: 429 and 941; or SEQ ID NO: 1453 and 1965); AD-20764 (SEQ ID NO: 452and 964; or SEQ ID NO: 1476 and 1988); AD-20783 (SEQ ID NO: 471 and 983;or SEQ ID NO: 1495 and 2007); AD-20278 (SEQ ID NOs: 2053 and 2064; orSEQ ID NOs: 2075 and 2086); AD-20279 (SEQ ID NOs: 2054 and 2065; or SEQID NOs: 2076 and 2087); AD-20280 (SEQ ID NOs: 2055 and 2066; or SEQ IDNOs: 2077 and 2088); AD-20281 (SEQ ID NOs: 2056 and 2067; or SEQ ID NOs:2078 and 2089); AD-20282 (SEQ ID NOs: 2057 and 2068; or SEQ ID NOs: 2079and 2090); AD-20283 (SEQ ID NOs: 2058 and 2069; or SEQ ID NOs: 2080 and2091); AD-20377 (SEQ ID NOs: 2059 and 2070; or SEQ ID NOs: 2081 and2092); AD-20570 (SEQ ID NOs: 2060 and 2071; or SEQ ID NOs: 2082 and2093); AD-20580 (SEQ ID NOs: 2061 and 2072; or SEQ ID NOs: 2083 and2094); AD-20597 (SEQ ID NOs: 2062 and 2073; or SEQ ID NOs: 2084 and2095); and AD-20598 (SEQ ID NOs: 2063 and 2074; or SEQ ID NOs: 2085 and2096). These example duplexes and the SEQ ID NOs for the specific sensestrand (SS) and antisense strand (AS) are provided herein with theirnucleotide sequence, e.g., as listed within Table 1, Table 2, Table 3,Table 3A, Table 8, Table 9A and Table 9B. Modified sequences (e.g.,sequences comprising one or more chemically modified base) of each ofthe compositions above and each of the RNAi agents in Tables 1, 2, 3,3A, 8, 9A and 9B are also contemplated as part of the disclosure.

Particular Compositions

In one embodiment, the present disclosure relates to particularcompositions comprising an RNAi agent comprising an antisense strand,wherein the antisense strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nucleotides from the antisensestrand of an RNAi agent to HSF1 selected from any one or more of thesequences in Tables 1, 2, 3, 3A, 8, 9A and 9B. In another embodiment,the present disclosure relates to a composition comprising an RNAi agentcomprising a sense strand and an antisense strand, wherein the antisensestrand comprises at least 15 contiguous nucleotides differing by 0, 1,2, or 3 nucleotides from the antisense strand of an RNAi agent to HSF1listed in any one or more of Tables 1, 2, 3, 3A, 8, 9A and 9B. Inanother embodiment, the present disclosure relates to a compositioncomprising an RNAi agent comprising a sense strand and an antisensestrand, wherein the sense strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nucleotides from the sense strandand the antisense strand comprises at least 15 contiguous nucleotidesdiffering by 0, 1, 2, or 3 nucleotides from the antisense strand of anRNAi agent to HSF1 listed immediately above. Particular duplexes includethose specific duplexes provided above and as listed in any one or moreof Table 1, Table 2, Table 3, Table 3A, Table 8, Table 9A and Table 9B.Additional modified sequences (e.g., sequences comprising one or moremodified base) of each of the compositions above are also contemplatedas part of the disclosure.

TABLE A1 SEQ ID NOs for Sense Strand (SS) and Antisense Strand (AS) forRNAi agents of the invention. RNAi agent SEQ ID NO: Strand: SS AS SS ASAD-20403 131 643 1155 1667 AD-20437 166 678 1190 1702 AD-20438 167 6791191 1703 AD-20439 168 680 1192 1704 AD-20487 169 681 1193 1705 AD-20489171 683 1195 1707 AD-20490 172 684 1196 1708 AD-20491 173 685 1197 1709AD-20548 234 746 1258 1770 AD-20560 269 781 1293 1805 AD-20562 271 7831295 1807 AD-20563 272 784 1296 1808 AD-20564 273 785 1297 1809 AD-20578285 797 1309 1821 AD-20626 290 802 1314 1826 AD-20627 291 803 1315 1827AD-20644 308 820 1332 1844 AD-20648 312 824 1336 1848 AD-20652 316 8281340 1852 AD-20660 324 836 1348 1860 AD-20694 377 889 1401 1913 AD-20707393 905 1417 1929 AD-20730 415 927 1439 1951 AD-20437.4 3218 3219AD-20487.7 3220 3221 AD-20489.2 3222 3223 AD-20560.4 3224 3225AD-37718.1 3226 3227 AD-37719.1 3242 3243 AD-37721.1 3228 3229AD-37722.1 3244 3245 AD-37724.1 3230 3231 AD-37725.1 3246 3247AD-37727.1 3232 3233 AD-37728.1 3248 3249 AD-37730.1 3234 3235AD-37733.1 3236 3237 AD-37736.1 3238 3239 AD-37740.1 3240 3241AD-36969.2 3250 3251 AD-30071.2 3252 3253 AD-36970.2 3254 3255AD-37739.1 3256 3257 AD-37731.1 3258 3259 AD-37734.1 3260 3261AD-37737.1 3262 3263 AD-37741.1 3264 3265 AD-37720.1 3266 3267AD-37723.1 3268 3269 AD-37726.1 3270 3271 AD-37729.1 3272 3273AD-37732.1 3274 3275 AD-37735.1 3276 3277 AD-37738.1 3278 3279AD-37742.1 3280 3281 AD-20303 30 542 1054 1566 AD-20313 40 552 1064 1576AD-20315 42 554 1066 1578 AD-20348 56 568 1080 1592 AD-20362 70 582 10941606 AD-20364 72 584 1096 1608 AD-20365 73 585 1097 1609 AD-20366 74 5861098 1610 AD-20373 81 593 1105 1617 AD-20376 84 596 1108 1620 AD-2037885 597 1109 1621 AD-20386 93 605 1117 1629 AD-20389 117 629 1141 1653AD-20391 119 631 1143 1655 AD-20392 120 632 1144 1656 AD-20397 125 6371149 1661 AD-20398 126 638 1150 1662 AD-20399 127 639 1151 1663 AD-20401129 641 1153 1665 AD-20402 130 642 1154 1666 AD-20404 132 644 1156 1668AD-20406 136 648 1160 1672 AD-20407 137 649 1161 1673 AD-20408 138 6501162 1674 AD-20409 139 651 1163 1675 AD-20410 140 652 1164 1676 AD-20411141 653 1165 1677 AD-20413 2042 2043 2046 2047 AD-20422 151 663 11751687 AD-20428 157 669 1181 1693 AD-20434 163 675 1187 1699 AD-20435 164676 1188 1700 AD-20488 170 682 1194 1706 AD-20493 175 687 1199 1711AD-20495 177 689 1201 1713 AD-20502 184 696 1208 1720 AD-20507 189 7011213 1725 AD-20513 195 707 1219 1731 AD-20527 209 721 1233 1745 AD-20535217 729 1241 1753 AD-20544 230 742 1254 1766 AD-20545 231 743 1255 1767AD-20546 232 744 1256 1768 AD-20547 233 745 1257 1769 AD-20549 235 7471259 1771 AD-20552 238 750 1262 1774 AD-20555 241 753 1265 1777 AD-20556242 754 1266 1778 AD-20557 243 755 1267 1779 AD-20558 267 779 1291 1803AD-20561 270 782 1294 1806 AD-20565 274 786 1298 1810 AD-20566 275 7871299 1811 AD-20572 280 792 1304 1816 AD-20574 2044 2045 2048 2049AD-20575 282 794 1306 1818 AD-20577 284 796 1308 1820 AD-20579 286 7981310 1822 AD-20625 289 801 1313 1825 AD-20633 297 809 1321 1833 AD-20634298 810 1322 1834 AD-20640 304 816 1328 1840 AD-20646 310 822 1334 1846AD-20650 314 826 1338 1850 AD-20653 317 829 1341 1853 AD-20661 325 8371349 1861 AD-20671 337 849 1361 1873 AD-20693 376 888 1400 1912 AD-20700383 895 1407 1919 AD-20702 385 897 1409 1921 AD-20709 394 906 1418 1930AD-20710 395 907 1419 1931 AD-20714 399 911 1423 1935 AD-20716 401 9131425 1937 AD-20728 413 925 1437 1949 AD-20741 429 941 1453 1965 AD-20764452 964 1476 1988 AD-20783 471 983 1495 2007 AD-20278 2053 2064 20752086 AD-20279 2054 2065 2076 2087 AD-20280 2055 2066 2077 2088 AD-202812056 2067 2078 2089 AD-20282 2057 2068 2079 2090 AD-20283 2058 2069 20802091 AD-20377 2059 2070 2081 2092 AD-20570 2060 2071 2082 2093 AD-205802061 2072 2083 2094 AD-20597 2062 2073 2084 2095 AD-20598 2063 2074 20852096

An RNAi Agent Comprising an Antisense Strand of an RNAi Agent DescribedHerein.

In one particular specific embodiment, the present disclosure relates toa composition comprising an RNAi agent comprising an antisense strand,wherein the antisense strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nucleotides from the antisensestrand of an RNAi agent to HSF1 selected from those antisense strands inthe specific duplexes provided above and as listed in Table 1, Table 2,Table 3, Table 3A, Table 8, Table 9A, and Table 9B.

Various particular specific embodiments of this embodiment are describedbelow.

In one embodiment, the composition further comprises a second RNAi agentto HSF1. In various embodiments, the second RNAi agent is physicallyseparate from the first, or the two are physically connected (e.g.,covalently linked or otherwise conjugated).

In one embodiment, the antisense strand is about 30 or fewer nucleotidesin length.

In one embodiment, the antisense strand forms a duplex region with asense strand, wherein the duplex region is about 15 to 30 nucleotidepairs in length.

In one embodiment, the antisense strand is about 15 to about 30nucleotides in length, including about 19 to about 23 nucleotides inlength. In one embodiment, the antisense strand has at least the lengthselected from about 15 nucleotides, about 16 nucleotides, about 17nucleotides, about 18 nucleotides, about 19 nucleotides, about 20nucleotides, about 21 nucleotides, about 22 nucleotides, about 23nucleotides, about 24 nucleotides, about 25 nucleotides, about 26nucleotides, about 27 nucleotides, about 28 nucleotides, about 29nucleotides and 30 nucleotides.

In one embodiment, the RNAi agent comprises a modification that causesthe RNAi agent to have increased stability in a biological sample orenvironment.

In one embodiment, the RNAi agent comprises at least one sugar backbonemodification (e.g., phosphorothioate linkage) or at least one2′-modified nucleotide.

In one embodiment, the RNAi agent comprises: at least one5′-uridine-adenine-3′ (5′-ua-3′) dinucleotide, wherein the uridine is a2′-modified nucleotide; at least one 5′-uridine-guanine-3′ (5′-ug-3′)dinucleotide, wherein the 5′-uridine is a 2′-modified nucleotide; atleast one 5′-cytidine-adenine-3′ (5′-ca-3′) dinucleotide, wherein the5′-cytidine is a 2′-modified nucleotide; or at least one5′-uridine-uridine-3′ (5′-uu-3′) dinucleotide, wherein the 5′-uridine isa 2′-modified nucleotide. These dinucleotide motifs are particularlyprone to serum nuclease degradation (e.g. RNase A). Chemicalmodification at the 2′-position of the first pyrimidine nucleotide inthe motif prevents or slows down such cleavage. This modification recipeis also known under the term ‘endo light’.

In one embodiment, the RNAi agent comprises a 2′-modification selectedfrom the group consisting of: 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl,2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP),2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl(2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), and2′-O-N-methylacetamido (2′-O-NMA). In one embodiment, all pyrimidines(uridine and cytidine) are 2′ O-methyl-modified nucleosides.

In one embodiment, the RNAi agent comprises at least one blunt end.

In one embodiment, the RNAi agent comprises an overhang having 1 nt to 4nt unpaired.

In one embodiment, the RNAi agent comprises an overhang at the 3′-end ofthe antisense strand of the RNAi agent.

In one embodiment, the RNAi agent is ligated to one or more diagnosticcompound, reporter group, cross-linking agent, nuclease-resistanceconferring moiety, natural or unusual nucleobase, lipophilic molecule,cholesterol, lipid, lectin, steroid, uvaol, hecigenin, diosgenin,terpene, triterpene, sarsasapogenin, Friedelin,epifriedelanol-derivatized lithocholic acid, vitamin, carbohydrate,dextran, pullulan, chitin, chitosan, synthetic carbohydrate, oligolactate 15-mer, natural polymer, low- or medium-molecular weightpolymer, inulin, cyclodextrin, hyaluronic acid, protein, protein-bindingagent, integrin-targeting molecule, polycationic, peptide, polyamine,peptide mimic, and/or transferrin.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 60% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 70% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 75% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 80% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 90% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 95% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 99% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi has an EC50 of no more than about 0.1 nM.EC50 is effective concentration to reduce gene expression by 50%.

In one embodiment, the RNAi has an EC50 of no more than about 0.01 nM.

In one embodiment, the RNAi has an EC50 of no more than about 0.001 nM.

An RNAi Agent Comprising a Sense and Antisense Strand of an RNAiDescribed Herein.

In one particular specific embodiment, the present disclosure relates toa composition comprising an RNAi agent comprising a sense strand and anantisense strand, wherein the sense strand and antisense strand compriseat least 15 contiguous nucleotides differing by 0, 1, 2, or 3nucleotides, from the sense and antisense strand, respectively, of anRNAi agent to HSF1 selected from the specific duplexes provided aboveand as listed in Table 1, Table 2, Table 3, Table 3A, Table 8, Table 9A,and Table 9B.

Various particular specific embodiments of this embodiment are describedbelow.

In one embodiment, the composition comprises a second RNAi agent toHSF1. In various embodiments, the second RNAi agent is physicallyseparate from the first, or the two are physically connected (e.g.,chemically linked or otherwise conjugated).

In one embodiment, the antisense strand is about 30 or fewer nucleotidesin length.

In one embodiment, the sense strand and the antisense strand form aduplex region about 15 to about 30 nucleotide pairs in length.

In one embodiment, the antisense strand and the sense strand are bothabout 19 to about 23 nt in length.

In one embodiment, the RNAi agent comprises a modification that causesthe RNAi agent to have increased stability in a biological sample orenvironment.

In one embodiment, the RNAi agent comprises a modified sugar backbonesuch as, e.g., a phosphorothioate linkage, or comprises a 2′-modifiednucleotide.

In one embodiment, the RNAi agent comprises: at least one5′-uridine-adenine-3′ (5′-ua-3′) dinucleotide, wherein the uridine is a2′-modified nucleotide; at least one 5′-uridine-guanine-3′ (5′-ug-3′)dinucleotide, wherein the 5′-uridine is a 2′-modified nucleotide; atleast 5′-cytidine-adenine-3′ (5′-ca-3′) dinucleotide, wherein the5′-cytidine is a 2′-modified nucleotide; or at least one5′-uridine-uridine-3′ (5′-uu-3′) dinucleotide, wherein the 5′-uridine isa 2′-modified nucleotide.

In one embodiment, the RNAi agent comprises a 2′-modification selectedfrom the group consisting of: 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl,2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP),2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl(2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), and2′-O—N-methylacetamido (2′-O-NMA). In one embodiment, all pyrimidines(uridine and cytidine) are 2′ O-methyl-modified nucleosides.

In one embodiment, the RNAi agent comprises at least one blunt end.

In one embodiment, the RNAi agent comprises an overhang having 1 to 4 ntunpaired.

In one embodiment, the RNAi agent comprises an overhang at the 3′-end ofthe antisense strand of the RNAi agent.

In one embodiment, the RNAi agent is ligated to one or more diagnosticcompound, reporter group, cross-linking agent, nuclease-resistanceconferring moiety, natural or unusual nucleobase, lipophilic molecule,cholesterol, lipid, lectin, steroid, uvaol, hecigenin, diosgenin,terpene, triterpene, sarsasapogenin, Friedelin,epifriedelanol-derivatized lithocholic acid, vitamin, carbohydrate,dextran, pullulan, chitin, chitosan, synthetic carbohydrate, oligolactate 15-mer, natural polymer, low- or medium-molecular weightpolymer, inulin, cyclodextrin, hyaluronic acid, protein, protein-bindingagent, integrin-targeting molecule, polycationic, peptide, polyamine,peptide mimic, and/or transferrin.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 60% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 70% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 80% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 90% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 95% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi agent is capable of inhibiting expression ofHSF1 by at least about 99% in WI-38 and/or HeLa cells in vitro.

In one embodiment, the RNAi has an EC50 of no more than about 0.1 nM.

In one embodiment, the RNAi has an EC50 of no more than about 0.01 nM.

In one embodiment, the RNAi has an EC50 of no more than about 0.001 nM.

A Method of Treatment Using an RNAi Agent Comprising a RNAi AgentDescribed Herein.

In one particular specific embodiment, the present disclosure relates toa method of treating a HSF1-related disease in an individual, comprisingthe step of administering to the individual a therapeutically effectiveamount of a composition comprising an RNAi agent comprising an antisensestrand, wherein the antisense strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nucleotides from the antisensestrand of an RNAi agent to HSF1 selected from those specific duplexesprovided above and as listed in Table 1, Table 2, Table 3, Table 3A,Table 8, Table 9A and Table 9B. In one embodiment, the RNAi agent toHSF1 comprises an antisense strand duplexed with a sense strand, whereinthe sense and antisense strands are selected from one or more of thesequences provided in Table 1, Table 2, Table 3, Table 3A, Table 8,Table 9A or Table 9.

Various particular specific embodiments of this embodiment are describedbelow.

In one embodiment, the HSF1-related disease is proliferative diseasesuch as, e.g., a cancer, or is an autoimmune disease, or is a viraldisease.

In one embodiment, the HSF1-related disease is cancer selected from thelist of bladder, bone, breast, cervical, colon, colorectal, endometrial,fibrosarcoma, gastric, haematopoietic, intestine, kidney, liver, lung,lymphoma, neuroectodermal, neuroblastoma, Ewing's sarcoma, osteosarcoma,ovary, pancreas, pleura, prostate, skin, squamous cell, stomach, andtesticular cancers, leukemia, promyelocytic leukemia, and Hodgkin'sdisease.

In one embodiment, the method further comprises the step ofadministering an additional cancer treatment.

In one embodiment, the method further comprises the step ofadministering an additional cancer treatment selected from the list ofactinomycin D, an inhibitor of HSP90 (heat shock protein 90), 17-AAG(tanespimycin), 17-DMAG (alvespimycin), IPI-504 (retaspimycin), IPI-493,SNX-5422 mesylate, AUY922, BIB021 CNF-2024, BIIB028, STA-9090, KW-2478,ATI3387, XL888, HSP990, MPC-3100, ABI-010 (as reviewed in Kim et al.2009 Curr. Topics in Med. Chem. 9: 1479-1492), or2-chlorodeoxyadenosine, 5-azacitidine, 5-fluoro-29-deoxyuridine,5-fluorouracil, 6-mercaptopurine, 6-thioguanine, 7-hydroxystaurosporine,13-cis-retinoic acid, a goserlin implant, alemtuzumab, alitretinoin,all-trans retinoic acid, alpha interferon, altretamine, amifostine,aminoglutethimide, anagrelide, anastrozole, arabinosylcytosine, arsenictrioxide, asparaginase, bacillus calmette-guerin, bendamustine,bevacizumab, bexarotene, bicalutamide, bleomycin, bortezomib, busulfan,camptothecin, capecitabine, carboplatin, carmustine, cetuximab,chlorambucil, cisplatin, cladribine, colcemid, Cycloheximide,cyclophosphamide, cytarabine, cytosine arabinoside (Ara-C), dacarbazine,dactinomycin, dasatinib, daunorubicin liposomal, daunorubicin,decitabine, denileukin diftitox, dexamethazone, docetaxel, doxorubicin,edelfosine, ehlorambucil, epipodophyllotoxin, epirubicin, erlotinib,estramustine, etoposide, everolimus, exemestane, fenretinide,finasteride, flavopiridol, floxuridine, fludarabine, fluorouracil,fluoxymesterone, flutamide, fulvestrant, gefitinib, gemeitabine,gemtuzumab ozogamicin, goserelin, hexamethylmelamine, hydrocortisone,hydroxyurea, ibritumomab tiuxetan, ibtritumomab, idarubicin, ifosfamide,imatinib, imidazole carboxamide, interleukin-11, interleukin-2,irinotecan, ixabepilone, lapatinib, L-asparaginase, lenalidomide,letrozole, leukovorin, leuprolide, mechlorethamine, megestrol,melphalan, mercaptopurine, methotrexate, methylprednisolone,mitixantrone, mitomycin, mitoxantrone, nelarabine, nitrogen mustard,octreotide, oxaliplatin, paclitaxel, paclitaxel-albumin formulations,paclitaxel-protein formulations, pamidronate, panitumumab, pemetrexed,pentostatin, phenylalanine mustard, pirubicin, prednisolone, prednisone,procarbazine, Puromycin, raloxifene, rituxan, rubidomycin, sargramostim,sorafenib, staurosporine, steroids, streptozocin, sunitinib, tamoxifen,Taxol, tegafur, temozolomide, temsirolimus, teniposide, thalidomide,thiophosphoamide, thiotepa, topotecan, toremifene, tositumomab,trastuzumab, tretinoin, UFT, vinblastine, vincristine, vinorelbine,vorinostat, and/or zoledronic acid. A RNAi agent to HSF1 can be used inconjunction with any additional treatment disclosed herein, asappropriate for the disease, optionally, in further conjunction with oneor more additional RNAi agents to HSF1.

In one embodiment, the HSF1-related disease is a viral disease.

In one embodiment, the HSF1-related disease is a viral disease selectedfrom the list of viral diseases mediated in whole or in part byadenovirus, herpes simplex virus, human cytomegalovirus, HTLV-1, SV40,polyoma virus, HIV, and/or Epstein-Barr virus.

In one embodiment, the method further comprises the step ofadministering an additional viral disease treatment.

In one embodiment, the method further comprises the step ofadministering an additional viral disease treatment selected from thelist of Abacavir, Aciclovir, acyclovir (acycloguanosine), Adefovir,Amantadine, Ampligen, Amprenavir, Arbidol, Atazanavir, Atripla,bevirimat, Boceprevir, broad spectrum inhibitor, Cidofovir, Combivir,Darunavir, Delavirdine, Didanosine, Docosanol, Edoxudine, Efavirenz,Emtricitabine, Enfuvirtide, Entecavir, Entry inhibitors, Entry or fusioninhibitor, Famciclovir, Fomivirsen, Fosamprenavir, Foscarnet, Fosfonet,Fusion inhibitor, Ganciclovir, Ibacitabine, Idoxuridine, Imiquimod,Imunovir, Indinavir, Inosine, Integrase inhibitor, Integrase inhibitor,Interferon, Interferon type I, Interferon type II, Interferon type III,Lamivudine, Lopinavir, Loviride, Maraviroc, Maturation inhibitor,Moroxydine, Nelfinavir, Nevirapine, Nexavir, Non-nucleoside reversetranscriptase inhibitor, NOV-205, Nucleoside analogues, Nucleotideanalog reverse transcriptase inhibitor, Oseltamivir (Tamiflu),Peginterferon alfa-2a, Penciclovir, Peramivir, Pleconaril,Podophyllotoxin, Protease inhibitor, Protease inhibitor, Raltegravir,Raltegravir, Reverse transcriptase inhibitor, Reverse transcriptaseinhibitor, Ribavirin, Rimantadine, Ritonavir, Saquinavir, Saquinavir,Stavudine, Synergistic enhancer (antiretroviral), Tenofovir, Tenofovirdisoproxil, Tipranavir, Trifluridine, Trizivir, Tromantadine, Truvada,Valaciclovir (Valtrex), Valganciclovir, Vicriviroc, Vidarabine,Viramidine, Zalcitabine, Zanamivir (Relenza), and Zidovudine. It will beunderstood that references to any additional treatment (e.g., viraldisease treatment or cancer treatment or autoimmune disease treatment,etc.) are meant to also include the pharmaceutically acceptable salts ofany of the active substances. If active substances comprised bycomponents (a) and/or (b) have, for example, at least one basic center,they can form acid addition salts. Corresponding acid addition salts canalso be formed having, if desired, an additionally present basic center.Active substances having an acid group, e.g., COOH, can form salts withbases. The active substances comprised in components (a) and/or (b) or apharmaceutically acceptable salts thereof may also be used in form of ahydrate or include other solvents used for crystallization.

In one embodiment, the HSF1-related disease is an autoimmune disease.

In one embodiment, the HSF1-related disease is lupus or rheumatoidarthritis.

In one embodiment, the composition comprises a second RNAi agent toHSF1. In various embodiments, the second RNAi agent is physicallydistinct from the first, or the two are physically connected (e.g.,linked or conjugated).

A Method of Inhibiting the Expression of HSF1, Using an RNAi Comprisingan RNAi Agent Described Herein.

In one particular specific embodiment, the present disclosure relates toa method of inhibiting the expression of HSF1 in an individual,comprising the step of administering to the individual a therapeuticallyeffective amount of a composition comprising an RNAi agent of thedisclosure. In one embodiment, the RNAi comprises a sense strand and anantisense strand, wherein the antisense strand comprises at least 15contiguous nucleotides differing by 0, 1, 2, or 3 nucleotides from theantisense strand of an RNAi agent to HSF1 selected from those specificduplexes provided above and as listed in Table 1, Table 2, Table 3,Table 3A, Table 8, Table 9A, and Table 9B.

Various particular specific embodiments of this embodiment are describedbelow.

In one embodiment, the individual is afflicted with or susceptible to anHSF1-related disease.

In one embodiment, the HSF1-related disease is proliferative disease,such as, e.g., a cancer.

In one embodiment, the HSF1-related disease is cancer selected from thelist of bladder, bone, breast, cervical, colon, colorectal, endometrial,fibrosarcoma, gastric, haematopoietic, intestine, kidney, liver, lung,lymphoma, neuroectodermal, neuroblastoma, Ewing's sarcoma, osteosarcoma,ovary, pancreas, pleura, prostate, skin, squamous cell, stomach, andtesticular cancers, leukemia, promyelocytic leukemia, and Hodgkin'sdisease.

In one embodiment, the method further comprises the step ofadministering an additional cancer treatment.

In one embodiment, the method further comprises the step ofadministering an additional cancer treatment selected from the list ofany cancer treatment listed herein, or known in the art.

In one embodiment, the HSF1-related disease is a viral disease.

In one embodiment, the HSF1-related disease is a viral disease selectedfrom the list of viral diseases mediated in whole or in part byadenovirus, herpes simplex virus, human cytomegalovirus, HTLV-1, SV40,polyoma virus, HIV, and/or Epstein-Barr virus.

In one embodiment, the method further comprises the step ofadministering an additional viral disease treatment.

In one embodiment, the method further comprises the step ofadministering an additional viral disease treatment selected from thelist of any viral disease treatment listed herein.

In one embodiment, the HSF1-related disease is an autoimmune disease.

In one embodiment, the HSF1-related disease is lupus or rheumatoidarthritis.

In one embodiment, the composition further comprises a second RNAi agentto HSF1. In various embodiments, the second RNAi agent is physicallydistinct from the first, or the two are physically connected (e.g.,linked or conjugated).

Other Embodiments

Various particular specific embodiments of this disclosure are describedbelow.

In one embodiment, the disclosure pertains to a composition according toany of the above embodiments, for use in a method of treating aHSF1-related disease in an individual, the method comprising the step ofadministering to the individual a therapeutically effective amount of acomposition according to any of the claims.

Various particular specific embodiments of this embodiment are describedbelow.

In one embodiment, the disclosure pertains to the composition accordingto any of the above embodiments, for use in a method of inhibiting theexpression of HSF1 in an individual, the method comprising the step ofadministering to the individual a therapeutically effective amount of acomposition according to any of the above embodiments.

One embodiment of the disclosure is the use of a composition accordingto any of the above embodiments, in the manufacture of a medicament fortreatment of an HSF1-related disease.

In one embodiment, the HSF1-related disease is selected from cancer,viral disease or autoimmune disease.

In one embodiment, the disclosure pertains to the composition of any ofthe above embodiments, for use in the treatment of an HSF1-relateddisease.

In one embodiment, the HSF1-related disease is selected from cancer,viral disease or autoimmune disease.

In one embodiment, the disclosure relates to a method of inhibiting theexpression of HSF1 in an cell, comprising the step of introducing intothe cell a composition comprising an RNAi agent comprising an antisensestrand, wherein the antisense strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nucleotides from the antisensestrand of an RNAi agent to HSF1 selected from the HSF1 siRNAs disclosedherein.

In one embodiment, the disclosure relates to a method of inhibiting theexpression of HSF1 in an cell, comprising the step of introducing intothe cell a composition comprising an RNAi agent comprising a sensestrand and an antisense strand, wherein the antisense strand comprisesat least 15 contiguous nucleotides differing by 0, 1, 2, or 3nucleotides from the antisense strand, and the sense strand comprises atleast 15 contiguous nucleotides differing by 0, 1, 2, or 3 nucleotidesfrom the sense strand of an RNAi agent to HSF1 selected from the HSF1siRNAs disclosed herein.

Heat Shock Factor 1 (HSF1)

By “HSF1” is meant the gene or protein heat shock factor 1, or heatshock transcription factor 1 (HSTF1). HSF1 is the master regulator ofthe heat shock response, in which multiple genes are induced in responseto temperature increase and other stresses. HSF1 has been designatedHGNC ID HGNC:5224, on Chromosome 8q24.3. It (including homologues) isalso identified as: GenelD: 3297; RefSeq IDs NM_(—)005526; AccNo.M64673; Mouse Genome Database ID MGI:96238; Rat Genome Database IDRGD:620913; Entrez Gene ID 3297; CCDS IDs CCDS6419.1; Pubmed IDs1871105; Ensembl ID ENSG00000185122; OMIM ID (NCBI) 140580; UCSC ID(UCSC) uc003zbt.2; and/or UniProt ID (mapped data supplied by UniProt)Q00613.

The amino acid sequence of human HSF1 is provided as SEQ ID NO 2050.:

(SEQ ID NO: 2050) MDLPVGPGAAGPSNVPAFLTKLWTLVSDPDTDALICWSPSGNSFHVFDQGQFAKEVLPKYFKHNNMASFVRQLNMYGFRKVVHIEQGGLVKPERDDTEFQHPCFLRGQEQLLENIKRKVTSVSTLKSEDIKIRQDSVTKLLTDVQLMKGKQECMDSKLLAMKHENEALWREVASLRQKHAQQQKVVNKLIQFLISLVQSNRILGVKRKIPLMLNDSGSAHSMPKYSRQFSLEHVHGSGPYSAPSPAYSSSSLYAPDAVASSGPIISDITELAPASPMASPGGSIDERPLSSSPLVRVKEEPPSPPQSPRVEEASPGRPSSVDTLLSPTALIDSILRESEPAPASVTALTDARGHTDTEGRPPSPPPTSTPEKCLSVACLDKNELSDHLDAMDSNLDNLQTMLSSHGFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPRPPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAEDPTISLLTGSEPPKAKDPTVS

The functional domains of HSF1 have been delineated by mutagenesis. Asequence near the N terminus forms the DNA binding domain (numberedapproximately aa 13-121; or aa 16-120, Shi et al. Adjacent to this is ahydrophobic region comprising three “leucine zippers” that mediatemonomerization and trimerization (numbered approximately aa 126-217; or137-212; or 137-203. A fourth hydrophobic patch or leucine zipper liesat approximately aa 378-407; this region is involved in negativeregulation under non-stress conditions. The central part of the moleculecontains a region that regulates the activity of transcriptionalactivation domains in response to stress. Sequences within theregulatory domain undergo specific phosphorylation and dephosphorylationin response to stress. This regulatory domain, which is rich in serinesand prolines, lies at approximately aa 221-310, or 201-370. TheC-terminal portion of HSF1 contains the main transcriptional activationregions; this comprises the 100 most C-terminal amino acids, or aa395-503, or can perhaps be reduced to AD1 at aa 401-420 (Newton et al.).These domains are described in, inter alia, Green et al. 1995 Mol. Cell.Biol. 15: 3354-3362; and Shi et al. 1995 Mol. Cell. Biol. 15: 4309-4318.The HSF1 RNAi agent of the present disclosure can interact with aspecific functional domain or domains of HSF1.

In various embodiments, the RNAi agents of the present disclosurespecifically bind to HSF1 mRNA, in a sequence corresponding to afunctional domain, e.g., in a sequence near the N terminus that formsthe DNA binding domain; in the 4/3 hydrophobic repeat or “leucinezipper” that mediates trimerization; in the first, second, third orfourth leucine zipper; in the central part of the molecule that containsseveral elements that maintain HSF1 in its latent form, or that regulatethe activity of transcriptional activation domains in response tostress; in sequences within the regulatory domain that undergo specificphosphorylation and dephosphorylation in response to stress; in theC-terminal portion of HSF1 that contains the main transcriptionalactivation regions; in the arrays of amphipathic alpha-helical residuesin the amino-terminal domain of HSF family proteins that interact toform coiled coils; and/or in the fourth region of amphipathicalpha-helix in the carboxyl-terminal domain. In other embodiments, theRNAi agents of the present disclosure bind to the 5′ or 3′ UTR[untranslated region(s)].

In various embodiments, the RNAi agents of the present disclosure bindto HSF1 mRNA, but not in a sequence corresponding to a functionaldomain, e.g., not in a sequence near the N terminus that forms the DNAbinding domain; not the 4/3 hydrophobic repeat or “leucine zipper” thatmediates trimerization; not the first, second, third or fourth leucinezipper; not the central part of the molecule that contains severalelements that maintain HSF1 in its latent form, or that regulate theactivity of transcriptional activation domains in response to stress;not in sequences within the regulatory domain that undergo specificphosphorylation and dephosphorylation in response to stress; not in theC-terminal portion of HSF1 that contains the main transcriptionalactivation regions; not in the arrays of amphipathic alpha-helicalresidues in the amino-terminal domain of HSF family proteins thatinteract to form coiled coils; not in the fourth region of amphipathicalpha-helix in the carboxyl-terminal domain; or not in the 5′ or 3′UTRs. In another embodiment, the RNAi agents of the present disclosurebind to the HSF1 mRNA, but not in sequence spanning nt 322 to 340downstream of the gene transcription start site as described by Rossi etal. 2006 Cancer Res. 66:7678-7685.

HSF1-Related Diseases

As used herein, the phrase a “HSF1-related disease” means one or more ofthe following: a proliferative disease, including, e.g., a cancer,wherein the cancer is selected from one or more of cancers of bladder,bone, breast, cervical, colon, colorectal, endometrial, fibrosarcoma,gastric, haematopoietic, intestine, kidney, liver, lung, lymphoma,neuroectodermal, neuroblastoma, Ewing's sarcoma, osteosarcoma, ovary,pancreas, pleura, prostate, skin, squamous cell, stomach, and testicularcancers, leukemia, promyelocytic leukemia, and Hodgkin's disease; aviral disease, wherein the viral disease is selected from one or more ofviral diseases mediated in whole or in part by adenovirus, herpessimplex virus, human cytomegalovirus, HTLV-1, SV40, polyoma virus, HIV,and/or Epstein-Barr virus; and an autoimmune disease, wherein theautoimmune disease is selected from one or more of lupus and arheumatoid arthritis.

HSF1 has been implicated in several diseases, including cancer and viraldiseases. HSF1 and other heat shock proteins (whose expression isincreased by HSF1) are over-expressed in, or have otherwise beenimplicated in bladder, bone, breast, cervical, colon, colorectal,endometrial, fibrosarcoma, gastric, haematopoietic, intestine, kidney,liver, lung, lymphoma, neuroectodermal, neuroblastoma, Ewing's sarcoma,osteosarcoma, ovary, pancreas, pleura, prostate, skin, squamous cell,stomach, and testicular cancers, leukemia (e.g., promyelocyticleukemia), and Hodgkin's disease. HSF1 is over-expressed in metastaticprostate carcinoma cell line PC-3M (as compared to the non-metastaticPC-3 line), and other prostate cancer cells.

The over-expression of HSF1 is correlated with an up-relation of heatshock protein HSP27. Hoang et al. 2000 Am. J. Pathol. 156: 857-864.HSP27 up-regulation is also associated with increased tumorigenicity andinvasiveness of some cancers, including colon, breast, promyelocyticleukemia, testicular and prostate. HSF1 also plays a functional role incancer cells under non-stress conditions; a dominant-negative HSF1alters DNA content in PC-3 cell populations and inhibits aneuploidy.Wang et al. 2004 J. Biol. Chem. 279: 32651-32659. Many tumor typescontain high concentrations of heat shock protein of the HSP27, HSP70,and HSP90 families, which are up-regulated by HSF1. Without wishing tobe bound by any particular theory, applicants note that it has beensuggested that heat shock proteins (HSP) may block the pathways ofapoptosis and permit malignant cells to arise despite the triggering ofapoptotic signals during transformation. HSP expression may also affordprotection of cancer cells from treatments such as chemotherapy andhyperthermia by thwarting the pro-apoptotic influence of thesemodalities. Tang et al. 2005 Cell Stress Chaperones 10: 46-58 andreferences therein. Rossi et al. also showed that decreasing HSF1 levelsincreased the sensitivity of uterine cervix carcinoma cells to cisplatinassociated with hyperthermia. Over-expression of heat shock proteins isalso associated with protection of cancer cells against doxorubicin andhyperthermia and other anti-cancer treatments. Helmbrecht et al. 2000Cell Prolif. 33: 341-365.

Over-expression of heat shock proteins is also associated with viralinfections, including those mediated by adenovirus, herpes simplexvirus, human cytomegalovirus, HTLV-1, SV40, polyoma virus, HIV,Epstein-Barr virus. High heat shock protein levels are also associatedwith autoimmune diseases, including lupus and rheumatoid arthritis.Inhibition of HSF1, e.g., via use of an anti-HSF1 RNAi agent, can thusbe an effective treatment against cancer, and viral and other diseases.Few HSP inhibitors are known, but they include quercetin, a flavonoidthat inhibits the HSF1. Zanini et al. 2007 J. Neurochem. 103:1344-354and references therein. Quercetin can thus be used as a positive controlfor RNAi agents that inhibit HSF1 in treating a viral disease or cancer.

HSF1 Gene Sequences in Various Species

The human HSF1 gene has been cloned, Rabindran et al. 1991 Proc. Natl.Acad. Sci USA 88: 6906-6910. Various sequences are available for humanHSF1, including Genbank identifier NM_(—)005526.2. The mouse (Musmusculus) HSF1 gene is, for example, Genbank id NM_(—)008296.2. Anothermouse HSF1 sequence is available as Acc. Number XM_(—)128055 (as used inYin et al. 2005 J. Mol. Cell. Card. 39: 681-689).

The Cynomolgus monkey (“Cyno”, or Macaca fascicularis) HSF1 sequence(SEQ ID NO: 2051), compared to the human sequence (SEQ ID NO: 2052), ispresented below in Table A2:

TABLE A2 humanGCGGCGGGAGCGCGCCCGTTGCAAGATGGCGGCGGCCATGCTGGGCCCCGGGGCTGTGTG cyno----------CGCGCCCGTTGCAAGATGGCGGCGGCAAAGCTGGGCCTTGGGGCTGGGGG          ************************** * ********  ******* * * humanTGCGCAGCGGGCGGCGGCGCGGCCCGGAAGGCTGGCGCGGCGACGGCGTTAGCCCGGCCC cynoGGCGCAGGGGGAGGCGGNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN  ****** ********                                                   Start-> humanTCGGCCCCTCTTTGCGGCCGCTCCCTCCGCCTATTCCCTCCTTGCTCGAG ATG GATCTGC cynoNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNCGAG ATG GATCTGC                                              ************** humanCCGTGGGCCCCGGCGCGGCGGGGCCCAGCAAC-GTCCCGGCCTTCCTGACCAAGCTGTGG cynoCCGTGGGCCCCGGTGCGGCGGGGCCCAGCAANCGTCCCGGCCTTCCTGACCAAGCTGTGG************* *****************  *************************** humanACCCTCGTGAGCGACCCGGACACCGACGCGCTCATCTGCTGGAGCCCGAGCGGGAACAGC cynoACCCTCGTGAGCGACCCGGACACCGACGCGCTCATCTGCTGGAGCCCGAGTGGGAACAGC************************************************** ********* humanTTCCACGTGTTCGACCAGGGCCAGTTTGCCAAGGAGGTGCTGCCCAAGTACTTCAAGCAC cynoTTCCATGTGTTCGACCAGGGCCAGTTTGCCAAGGAGGTGCTGCCCAAGTATTTCAAGCAC ************************************************* ********* humanAACAACATGGCCAGCTTCGTGCGGCAGCTCAACATGTATGGCTTCCGGAAAGTGGTCCAC cynoAACAACATGGCCAGCTTCGTGCGGCAGCTCAACATGTATGGTTTCCGGAAAGTGGTCCAC***************************************** ****************** humanATCGAGCAGGGCGGCCTGGTCAAGCCAGAGAGAGACGACACGGAGTTCCAGCACCCATGC cynoATCGAGCAGGGTGGCCTGGTCAAGCCAGAGAGAGACGACACGGAGTTCCAGCACCCGTGC ******************************************************* *** humanTTCCTGCGTGGCCAGGAGCAGCTCCTTGAGAACATCAA-GAGGAAAGTGACCAGTGTGTC cynoTTCCTGCGCGGCCAGGAGCAGCTCCTTGAGAACATCANAGAGGAAAGTGACCAGTGTGTC ************************************  ********************* humanCACCCTGAAGAGTGAAGACATAAAGATCCGCCAGGACAGCGTCACCAAGCTGCTGACGGA cynoCACCCTGAAGAGTGAAGACATAAAGATCCGTCAGGACAGTGTCACCAAGCTGCTGACGGA****************************** ******** ******************** humanCGTGCAGCTGATGAAGGGGAAGCAGGAGTGCATGGACTCCAAGCTCCTGGCCATGAAGCA cynoCGTGCAGCTGATGAAGGGGAAGCAGGAGTGCATGGACTCCAAGCTCCTGGCCATGAAGCA************************************************************ humanTGAGAATGAGGCTCTGTGGCGGGAGGTGGCCAGCCTTCGGCAGAAGCATGCCCAGCAACA cynoTGAGAATGAGGCTCTGTGGCGGGAGGTGGCCAGCCTTCGGCAGAAGCATGCCCAGCAACA************************************************************ humanGAAAGTCGTCAACAAGCTCATTCAGTTCCTGATCTCACTGGTGCAGTCAAACCGGATCCT cynoGAAAGTCGTCAACAAGCTCATTCAGTTCCTGATCTCACTGGTGCAGTCAAACCGGATCCT************************************************************ humanGGGGGTGAAGAGAAAGATCCCCCTGATGCTGAACGACAGTGGCTCAGCACATTCCATGCC cynoGGGGGTGAAGAGAAAGATCCCCCTGATGCTGAACGACAGTGGCTCAGCACATTCCATGCC************************************************************ humanCAAGTATAGCCGGCAGTTCTCCCTGGAGCACGTCCACGGCTCGGGCCCCTACTCGGCCCC cynoCAAGTATGGCCGGCAGTTCTCCCTGGAGCACGTCCACGGCTCGGGCCCCTACTCGGCCCC *********************************************************** humanCTCCCCAGCCTACAGCAGCTCCAGCCTCTACGCCCCTGATGCTGTGGCCAGCTCTGGACC cynoCTCCCCAGCCTACAGTAGCTCCAGCCTCTACGCCCCCGATTCTGTGGCCAACTCCGGACC*************** ******************** *** ********* *** ***** humanCATCATCTCCGACATCACCGAGCTGGCTCCTGCCAGCCCCATGGCCTCCCCCGGCGGGAG cynoCATCATCTCCGACATCACCGAGCTGGCTCCTGCCAGCCCCGTGGCCTCCCCTGGCGGGAG**************************************** ********** ******** humanCATAGACGAGAGGCCCCTATCCAGCAGCCCCCTGGTGCGTGTCAAGGAGGAGCCCCCCAG cynoCATAGACGAGAGGCCCCTGTCTAGCAGCCCCCTGGTGCGTGTCAAAGAGGAGCCCCCCAG****************** ** *********************** ************** humanCCCGCCTCAGAGCCCCCGGGTAGAGGAGGCGAGTCCCGGGCGCCCATCTTCCGTGGACAC cynoCCCGCCTCAGAGCCCCCGGGTAGAGGAGGCGAGTCCCGGGCGCCCATCTTCCGTGGACAC************************************************************ humanCCTCTTGTCCCCGACCGCCCTCATTGACTCCATCCTGCGGGAGAGTGAACCTGCCCCCGC cynoCCTCTTGTCCCCGACCGCCCTCATTGACTCCATCCTGCGGGAGAGTGAACCTACCCCCGC**************************************************** ******* humanCTCCGTCACAGCCCTCACGGACGCCAGGGGCCACACGGACACCGAGGGCCGGCCTCCCTC cynoCTCCGCCACAGCCCTCACCGATGCCAGGGGCCACACGGACACCGAGGGCCGGCCTCCCTC ***************** ** ************************************** humanCCCCCCGCCCACCTCCACCCCTGAAAAGTGCCTCAGCGTAGCCTGCCTGGACAAGAATGA cynoACCCCCGCCCACCTCCACCCCTGAAAAGTGCCTCAGCGTAGCCTGCCTGGACAAGAATGA *********************************************************** humanGCTCAGTGACCACTTGGATGCTATGGACTCCAACCTGGATAACCTGCAGACCATGCTGAG cynoGCTCAGTGATCACTTGGATGCTATGGACTCCAACCTGGACAACCTGCAGACCATGCTGAG ************************************** ******************** humanCAGCCACCGCTTCACCGTGGACACCAGTGCCCTGCTGGACCTGTTCAGCCCCTCGGTGAC cynoCAGCCACCGCTTCACCGTGGACACCACCGCCCTGCTGGACCTGTTCAGCCCCTCGGTGAC*************************** ******************************** humanCGTGCCCGACATGAGCCTGCCTGACCTTGACACCAGCCTGGCCAGTATCCAAGAGCTCCT cynoCGTGCCCGACATGAGCCTGCCTGACCTTGACACCAGCCTGGCTAGTATCCAAGAGCTCCT****************************************** ***************** humanGTCTCCCCAGGAGCCCCCCAGGCCTCCCGAGGCAGAGAACACCAGCCCGGATTCAGGGAA cynoGTCTCCCCAGGAGCCCTCCAGGCCTCCCGAGGCAGAGAACACCAGCCCGGATTCAGGGAA**************** ******************************************* humanGCAGCTGGTGCACTACACACCGCAGCCGCTGTTCCTGCTGGACCCCGGCTCCGTGGACAC cynoGCAGCTGGTGCACTACACAGCACAGCCACTGTTCCTGCTCGACCCCGGCTCCGTGGGCAC********************* ***** *********** **************** *** humanCGGGACCAACGACCTGCCGGTGCTGTTTGAGCTGGGAGAGGGCTCCTACTTCTCCGAAGG cynoCGGGAGCAGCGACTTGCCGGTGCTGTTTGAGCTGGGGGAGGGCTCCTACTTCTCCGAAGG ************ ********************** *********************** humanGGACGGCTTCGCCGAGGACCCCACCATCTCCCTGCTGACAGGCTCGGAGCCTCCCAAAGC cynoGGACGGCTTCGCAGAGGACCCCACCATCTCCCTGCTGACAGGCTCAGAGCCTCCCAAAGC************ ******************************** **************                   STOP human CAAGGACCCCACTGTCTCC TAGAGGCCCCGGAGGACCTGGGCCAGCCGCCCACCCCCACC cyno CAAGGACCCCACTGTCTCC TAGGCGCCCGGGAGGAGCTGGGCCAGCCGCCCACCCCCACC **********************  *********************************** humanCCCAGTGCAGGGCTGGTCTTGGGGAGGCAG-GGCAGCCTCGCGGTCTTGGGCACTGGTGG cynoCCCAGTGCAGGGCTGGCCTTGGGGAGGAAGAGGCAGCCTCGAGGTCCTGGGCACTGGTGG**************** ********** ** ********** **** ************* humanGTCGGCCGCCATAGCCCCAGTAGGACAAAC--GGGCTCGGGTCTGGGCAGCACCTCTGGT cynoGTTGGCCACCACAGCCCCAGTAGGACAAACAGGGGCTCAGGTCTGGGCAGCACCTCTGGT ** **** *********************  ****** ********************* humanCAGGAGGGTCACCCTGGCCTGCCAGTCTGCCTTCCCCCAACCCCGTGTCCTGTGGTTTGG cynoCAGGAGGGTCACCCCGGCCTCCCAGTCTGCCTTCCCCCAACCCCGTGTCCTGTGGTTTGG************** ***** *************************************** humanTTGGGGCTTCACAGCCACACCTGGACTGACCCTGCAGGTTGTTCATAGTCAGAATTGTAT cynoTTGGGGCTTCGTAGCCACACCTGGACTGACCCTGCAGGTTGTTCATAATCAGAATTGTAT**********  *********************************** ************ humanTTTGGATTTTTACACAACTGTCCCGTTCCCCGCTCCACAGAGATACACAGATATATACAC cynoTTTGGATTTTTACACAACTGTCCCATTCCCTGTTCCATAGAGATATACAGATATATACAC************************ ***** * **** ******* ************** humanACAG-TGGATGGACGGACAAGACAGGCAGAGATCTATAAACAGACAGGCTCTATGCTAAA cynoACAGGTGGATGGACGGACAAGACAGGCAGAGATCTATAAACAGACAG------------- ********************************************** human AAAAAAAAAAAA (SEQ IDNO: 2051) cyno ------------ (SEQ ID NO: 2052)

The start (ATG) and stop (TAG) of the human HSF1 sequence and putativestart and stop of the cyno HSF1 sequence are indicated in boldunderlined. N indicates that the nucleotide was not determined at thatposition in the sequencing experiment.

In one embodiment, the HSF1 RNAi agent of the present disclosurecomprises a sequence which is identical in the human, mouse and cynoHSF1 gene. This sequence identity facilitates animal testing prior tohuman testing.

In one embodiment, the HSF1 RNAi agent comprises a sequence which doesnot match that of any other gene. In one embodiment, the HSF1 RNAi agentcomprises a sequence which differs from all other known non-HSF1 genesby at least 0, 1, 2 or 3 nucleotides.

In one embodiment, the HSF1 RNAi agent comprises a sequence which isidentical to that in HSF2, HSF3 or HSF4. In one embodiment, the HSF1RNAi agent comprises a sequence which is not identical to any in HSF2,HSF3 or HSF4.

HSF1 RNAi Agent for Use in Treating Various HSF1-Related Diseases

In one embodiment, the HSF1 RNAi agent of the present disclosurecomprises a sequence disclosed herein and is administered to a patientin need thereof (e.g., a patient suffering from cancer and/or a viraldisease and/or autoimmune disease and/or HSF1-related disease). In oneembodiment, the HSF1 RNAi agent of the present disclosure isadministered to a patient in need thereof, along with one or moreadditional pharmaceutical agent appropriate for that disease. Forexample, a patient suffering from cancer can be administered apharmacologically effective amount of one or more HSF1 RNAi agent alongwith a pharmacologically effective amount of one or more of any cancertreatment listed herein, and/or any other cancer treatment known in theart.

A patient suffering from a viral disease can be administered one or moreRNAi agent to HSF1 and one or more additional viral disease treatment.This additional treatment can be selected from the list of any viraldisease treatment listed herein, and/or any anti-viral known in the art.

The patient can also be administered more than one RNAi agent to HSF1.

In the case of cancer, autoimmune and viral diseases, the RNAi agent(s)and additional disease treatment(s) can be administered in any order,simultaneously or sequentially, or in multiple doses over time.Administration of the RNAi agent and the additional treatment can be,for example, simultaneous, concurrent, separate or sequential.

Simultaneous administration may, e.g., take place in the form of onefixed combination with two or more active ingredients, or bysimultaneously administering two or more active ingredients that areformulated independently. Sequential use (administration) preferablymeans administration of one (or more) components of a combination at onetime point, other components at a different time point, that is, in achronically staggered manner, preferably such that the combination showsmore efficiency than the single compounds administered independently(especially showing synergism). Separate use (administration) preferablymeans administration of the components of the combination independentlyof each other at different time points, preferably meaning that thecomponents (a) and (b) are administered such that no overlap ofmeasurable blood levels of both compounds are present in an overlappingmanner (at the same time).

Also combinations of two or more of sequential, separate andsimultaneous administration are possible, preferably such that thecombination component-drugs show a joint therapeutic effect that exceedsthe effect found when the combination component-drugs are usedindependently at time intervals so large that no mutual effect on theirtherapeutic efficiency can be found, a synergistic effect beingespecially preferred.

The term “delay of progression” as used herein means administration ofthe combination to patients being in a pre-stage or in an early phase,of the first manifestation or a relapse of the disease to be treated, inwhich patients, e.g., a pre-form of the corresponding disease isdiagnosed or which patients are in a condition, e.g., during a medicaltreatment or a condition resulting from an accident, under which it islikely that a corresponding disease will develop.

“Jointly therapeutically active” or “joint therapeutic effect” meansthat the compounds may be given separately (in a chronically staggeredmanner, especially a sequence-specific manner) in such time intervalsthat they preferably, in the warm-blooded animal, especially human, tobe treated, still show a (preferably synergistic) interaction (jointtherapeutic effect). Whether this is the case, can inter alia bedetermined by following the blood levels, showing that both compoundsare present in the blood of the human to be treated at least duringcertain time intervals.

Definitions

For convenience, the meaning of certain terms and phrases used in thespecification, examples, and appended claims, are provided below. Ifthere is an apparent discrepancy between the usage of a term in otherparts of this specification and its definition provided in this section,the definition in this section shall prevail.

As used throughout this disclosure, articles such as “a” and “an” referto one or more than one (at least one) of the grammatical object of thearticle.

RNAi Agent

As used herein, the term “RNAi agent,” “RNAi agent to HSF1”, “siRNA toHSF1”, “HSF1 siRNA” and the like refer to an siRNA (short inhibitoryRNA), shRNA (short or small hairpin RNA), iRNA (interference RNA) agent,RNAi (RNA interference) agent, dsRNA (double-stranded RNA), microRNA,and the like, which specifically binds to the HSF1 gene. As used herein,the terms “iRNA” and “RNAi” refers to an agent that contains RNA, andwhich mediates the targeted cleavage of another RNA transcript via anRNA-induced silencing complex (RISC) pathway. In one embodiment, theRNAi agent is an oligonucleotide composition that activates the RISCcomplex/pathway. In another embodiment, the RNAi agent comprises anantisense strand sequence (antisense oligonucleotide). In oneembodiment, the RNAi comprises a single strand. This single-strandedRNAi agent oligonucleotide or polynucleotide can comprise the sense orantisense strand, as described by Sioud 2005 J. Mol. Biol.348:1079-1090, and references therein. Thus the disclosure encompassesRNAi agents with a single strand comprising either the sense orantisense strand of an RNAi agent described herein.

RNA interference is a post-transcriptional, targeted gene-silencingtechnique that uses double-stranded RNA (dsRNA) to degrade messenger RNA(mRNA) containing the same sequence as the dsRNA. The process of RNAioccurs when ribonuclease III (Dicer) cleaves the longer dsRNA intoshorter fragments called siRNAs. siRNAs (small interfering RNAs) aretypically about 21 to 23 nucleotides long and comprise about 19 basepair duplexes. The smaller RNA segments then mediate the degradation ofthe target mRNA. Dicer has also been implicated in the excision of 21-and 22-nucleotide small temporal RNAs (stRNAs) from precursor RNA ofconserved structure that are implicated in translational control.Hutvagner et al. 2001, Science, 293, 834. The RNAi response alsofeatures an endonuclease complex, commonly referred to as an RNA-inducedsilencing complex (RISC), which mediates cleavage of single-strandedmRNA complementary to the antisense strand of the siRNA. Cleavage of thetarget RNA takes place in the middle of the region complementary to theantisense strand of the siRNA duplex.

Kits for RNAi synthesis are commercially available, e.g., from NewEngland Biolabs and Ambion.

The use of the RNAi agent to HSF1 results in a decrease of HSF1activity, level and/or expression, e.g., a “knock-down” or “knock-out”of the target gene or target sequence.

A suitable RNAi agent can be selected by any process known in the art orconceivable by one of ordinary skill in the art. For example, theselection criteria can include one or more of the following steps:initial analysis of the HSF1 gene sequence and design of RNAi agents;this design can take into consideration sequence similarity acrossspecies (human, cynomolgus, mouse, etc.) and dissimilarity to other(non-HSF1) genes; screening of RNAi agents in vitro (e.g., at 10 nM inWI-38 cells); determination of EC50 in HeLa cells; determination ofviability of WI-38, HeLa and GTL16 cells treated with RNAi agents,wherein it is desired that the RNAi agent to HSF1 not inhibit theviability of these cells; testing with human PBMC (peripheral bloodmononuclear cells), e.g., to test levels of TNF-alpha to estimateimmunogenicity, wherein immunostimulatory sequences are less desired;testing in human whole blood assay, wherein fresh human blood is treatedwith an RNAi agent and cytokine/chemokine levels are determined [e.g.,TNF-alpha (tumor necrosis factor-alpha) and/or MCP1 (monocytechemotactic protein 1)], wherein Immunostimulatory sequences are lessdesired; determination of gene knockdown in vivo using Hep3Bsubcutaneous tumors in test animals; HSF1 target gene modulationanalysis, e.g., using a pharmacodynamic (PD) marker, for example, HSP70or HSP27, wherein HSF1 knockdown leads to a dose-dependent reduction ofHSP70 and HSP27 expression in A375 cells; and optimization of specificmodifications of the RNAi agents.

Targets and Sequences

As used herein, “target sequence” or “target gene” refer to a contiguousportion of the nucleotide sequence of an mRNA molecule formed during thetranscription of a gene, e.g., a HSF1 gene, including mRNA that is aproduct of RNA processing of a primary transcription product. The targetportion of the sequence will be at least long enough to serve as asubstrate for iRNA-directed cleavage at or near that portion. Forexample, the target sequence will generally be from about 9-36nucleotides (“nt”) in length, e.g., about 15-30 nucleotides in length,including all sub-ranges therebetween. As non-limiting examples, thetarget sequence can be from about 15-30 nt, about 15-26 nt, about 15-23nt, about 15-22 nt, about 15-21 nt, about 15-20 nt, about 15-19 nt,about 15-18 nt, about 15-17 nt, about 18-30 nt, about 18-26 nt, about18-23 nt, about 18-22 nt, about 18-21 nt, about 18-20 nt, about 19-30nt, about 19-26 nt, about 19-23 nt, about 19-22 nt, about 19-21 nt,about 19-20 nt, about 20-30 nt, about 20-26 nt, about 20-25 nt, about20-24 nt, about 20-23 nt, about 20-22 nt, about 20-21 nt, about 21-30nt, about 21-26 nt, about 21-25 nt, about 21-24 nt, about 21-23 nt, orabout 21-22 nt.

As used herein, the term “strand comprising a sequence” refers to anoligonucleotide comprising a chain of nucleotides that is described bythe sequence referred to using the standard nucleotide nomenclature.

As used herein, and unless otherwise indicated, the term “complementary”refers to the ability of an oligonucleotide or polynucleotide comprisinga first nucleotide sequence to hybridize and form a duplex structureunder certain conditions with an oligonucleotide or polynucleotidecomprising a second nucleotide sequence. Such conditions can, forexample, be stringent, e.g., 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA,50° C. or 70° C. for 12-16 hours followed by washing. Other conditions,such as physiologically relevant conditions as may be encountered insidean organism, can apply. The skilled person will be able to determine theset of conditions most appropriate for a test of complementarity of twosequences in accordance with the ultimate application of the hybridizednucleotides.

Complementary sequences within an iRNA, e.g., within a dsRNA asdescribed herein, include base-pairing of the oligonucleotide orpolynucleotide comprising a first nucleotide sequence to anoligonucleotide or polynucleotide comprising a second nucleotidesequence over the entire length of one or both sequences. Such sequencescan be referred to as “fully complementary” with respect to each otherherein. However, where a first sequence is referred to herein as“substantially complementary” with respect to a second sequence, the twosequences can be fully complementary, or they may form one or more, butgenerally not more than 5, 4, 3 or 2 mismatched base pairs uponhybridization for a duplex up to 30 base pairs, while retaining theability to hybridize under the conditions most relevant to theirultimate application, e.g., inhibition of gene expression via a RISCpathway. However, where two oligonucleotides are designed to form, uponhybridization, one or more single stranded overhangs, such overhangsshall not be regarded as mismatches with regard to the determination ofcomplementarity. For example, a dsRNA comprising one oligonucleotide 21nt in length and another oligonucleotide 23 nt in length, wherein thelonger oligonucleotide comprises a sequence of 21 nt that is fullycomplementary to the shorter oligonucleotide, may yet be referred to as“fully complementary” for the purposes herein.

“Complementary” sequences, as used herein, may also include, or beformed entirely from, non-Watson-Crick base pairs and/or base pairsformed from non-natural and modified nucleotides, in as far as the aboverequirements with respect to their ability to hybridize are fulfilled.Such non-Watson-Crick base pairs includes, but are not limited to, G:UWobble or Hoogstein base pairing.

The terms “complementary,” “fully complementary” and “substantiallycomplementary” herein may be used with respect to the base matchingbetween the sense strand and the antisense strand of a dsRNA, or betweenthe antisense strand of an iRNA agent and a target sequence, as will beunderstood from the context of their use.

As used herein, a polynucleotide that is “substantially complementary toat least part of” a messenger RNA (mRNA) refers to a polynucleotide thatis substantially complementary to a contiguous portion of the mRNA ofinterest (e.g., an mRNA encoding HSF1). For example, a polynucleotide iscomplementary to at least a part of a HSF1 mRNA if the sequence issubstantially complementary to a non-interrupted portion of an mRNAencoding HSF1.

Double-Stranded RNA

The term “double-stranded RNA” or “dsRNA,” as used herein, refers to aniRNA that includes an RNA molecule or complex of molecules having ahybridized duplex region that comprises two anti-parallel andsubstantially complementary nucleic acid strands, which will be referredto as having “sense” and “antisense” orientations with respect to atarget RNA. The antisense strand, with respect to the mRNA target, isalso called the “guide” strand, and the sense strand is also called the“passenger” strand. The passenger strand can include at least one ormore of the following: one or more extra nucleotides (e.g., a bulge or 1nt loop) compared to the other strand, a nick, a gap, etc., compared tothe other strand.

The duplex region can be of any length that permits specific degradationof a desired target RNA through a RISC pathway, but will typically rangefrom 9 to 36 base pairs in length, e.g., 15-30 base pairs in length.Considering a duplex between 9 and 36 base pairs, the duplex can be anylength in this range, for example, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,or 36 and any sub-range therebetween, including, but not limited to15-30 base pairs (“bp”), 15-26 bp, 15-23 bp, 15-22 bp, 15-21 bp, 15-20bp, 15-19 bp, 15-18 bp, 15-17 bp, 18-30 bp, 18-26 bp, 18-23 bp, 18-22bp, 18-21 bp, 18-20 bp, 19-30 bp, 19-26 bp, 19-23 bp, 19-22 bp, 19-21bp, 19-20 bp, 20-30 bp, 20-26 bp, 20-25 bp, 20-24 bp, 20-23 bp, 20-22bp, 20-21 bp, 21-30 bp, 21-26 bp, 21-25 bp, 21-24 bp, 21-23 bp, or 21-22bp. dsRNAs generated in the cell by processing with Dicer and similarenzymes are generally in the range of about 19-22 base pairs in length.One strand of the duplex region of a dsDNA comprises a sequence that issubstantially complementary to a region of a target RNA. The two strandsforming the duplex structure can be from a single RNA molecule having atleast one self-complementary region, or can be formed from two or moreseparate RNA molecules. Where the duplex region is formed from twostrands of a single molecule, the molecule can have a duplex regionseparated by a single stranded chain of nucleotides (herein referred toas a “hairpin loop”) between the 3′-end of one strand and the 5′-end ofthe respective other strand forming the duplex structure. The hairpinloop can comprise at least one unpaired nucleotide; in some embodimentsthe hairpin loop can comprise at least 3, at least 4, at least 5, atleast 6, at least 7, at least 8, at least 9, at least 10, at least 20,at least 23 or more unpaired nucleotides. Where the two substantiallycomplementary strands of a dsRNA are comprised by separate RNAmolecules, those molecules need not, but can be covalently connected.Where the two strands are connected covalently by means other than ahairpin loop, the connecting structure is referred to as a “linker.” Theterm “siRNA” is also used herein to refer to a dsRNA as described above.

In one aspect, an RNA interference agent includes a single stranded RNAthat interacts with a target RNA sequence to direct the cleavage of thetarget RNA. Without wishing to be bound by theory, long double strandedRNA introduced into plants and invertebrate cells is broken down intosiRNA by a Type III endonuclease known as Dicer (Sharp et al., GenesDev. 2001, 15:485). Dicer, a ribonuclease-III-like enzyme, processes thedsRNA into 19-23 base pair short interfering RNAs with characteristictwo base 3′ overhangs (Bernstein, et al., (2001) Nature 409:363). ThesiRNAs are then incorporated into an RNA-induced silencing complex(RISC) where one or more helicases unwind the siRNA duplex, enabling thecomplementary antisense strand to guide target recognition (Nykanen, etal., (2001) Cell 107:309). Upon binding to the appropriate target mRNA,one or more endonucleases within the RISC cleaves the target to inducesilencing (Elbashir, et al., (2001) Genes Dev. 15:188). Thus, in oneaspect the disclosure relates to a single stranded RNA that promotes theformation of a RISC complex to effect silencing of the target gene.

Down-Regulation of HSF1

As used herein, “down-regulates” refers to any statistically significantdecrease in a biological activity and/or expression of HSF1, includingfull blocking of the activity (i.e., complete inhibition) and/orexpression. For example, “down-regulation” can refer to a decrease of atleast about 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100% in HSF1 activityand/or expression.

As used herein, the term “inhibit” or “inhibiting” HSF1 refers to anystatistically significant decrease in biological activity and/orexpression of HSF1, including full blocking of the activity and/orexpression. For example, “inhibition” can refer to a decrease of atleast about 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100% in HSF1 activityand/or expression. As used herein, the term “inhibit” similarly refersto a significant decrease in activity and/or expression, while referringto any other biological agent or composition.

By “level”, it is meant that the HSF1 RNAi agent can interfere with thedetectable level of HSF1, e.g., the level of HSF1 mRNA or the level ofHSF1 protein.

By “activity,” it is meant that the HSF1 RNAi agent can alter any knownactivity of HSF1, as described herein or as known in the literature.

By “heat shock” (HS) and “heat shock response” (HSR) is meant thebiochemical response to environmental stress, such as elevatedtemperature. In the laboratory, experimental animals and cells can bemaintained at a “non-shock” temperature (37° C. or lower) and heat shockcan be induced at an elevated temperature (e.g., 40, 41, 42, 43, 44, or45 degrees C. or higher). Experimentally, heat shock is typicallyinduced at 42, 43 or 44 degrees C.

Heat shock is characterized by misfolding, denaturation and aggregationof various proteins; the induced heat shock proteins (HSP or HSPs)include chaperone proteins (chaperonins) and others which repair and/orremove these proteins. Genes induced during the heat shock responseinclude, inter alia, HSP90, HSP70 and HSP27. The heat shock response canalso be induced (or mimicked) by additional environmental conditions,such as oxidative stress, chemical stress, free radicals, ATP depletion,acidosis, heavy metals, alcohols, presence of antibiotics, inhibitors ofenergy metabolism, pathological conditions such as ischemia andreperfusion, inflammation, tissue damage, infection and mutant proteinsassociated with genetic diseases. Jolly et al. 2000 J. Natl. CancerInst. 92: 1564-1572; Dai et al. 2007 Cell 130: 1005-1018.

The RNAi Agent to HSF1.

In one embodiment, the disclosure pertains to a HSF1 RNAi agent or otherantisense nucleic acid complementary to a HSF1 gene (or portionthereof), or a recombinant expression vector encoding the antisensenucleic acid. As used herein, an “antisense” nucleic acid comprises anucleotide sequence complementary to a “sense” nucleic acid encoding theHSF1 protein (e.g., complementary to the coding strand of adouble-stranded DNA, complementary to an mRNA or complementary to thecoding strand of a HSF1 gene).

The use of antisense nucleic acids to down-modulate the expression of aparticular protein in a cell is well known in the art. An antisensenucleic acid comprises a sequence complementary to, and is capable ofhydrogen binding to, the coding strand of another nucleic acid (e.g., anmRNA). Antisense sequences complementary to an mRNA can be complementaryto the coding region, the 5′ or 3′ untranslated region of the mRNA,and/or a region bridging the coding and untranslated regions, and/orportions thereof Furthermore, an antisense nucleic acid can becomplementary to a regulatory region of the gene encoding the mRNA, forinstance a transcription or translation initiation sequence orregulatory element. Preferably, an antisense nucleic acid can becomplementary to a region preceding or spanning the initiation codon onthe coding strand or in the 3′ untranslated region of an mRNA.

Antisense nucleic acids can be designed according to the rules of Watsonand Crick base pairing. The antisense nucleic acid molecule can becomplementary to the entire coding region of HSF1 mRNA, but in at leastone embodiment is an oligonucleotide which is antisense to only aportion of the coding or non-coding region of HSF1 mRNA. For example,the antisense oligonucleotide can be complementary to the regionsurrounding the translation start site of HSF1 mRNA. An antisenseoligonucleotide can be, for example, about 5, about 10, about 15, about16, about 17, about 18, about 19, about 20, about 21, about 22, about23, about 24, about 25, about 30, about 35, about 40, about 45 or about50 nt in length, or 5, 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,30, 35, 40, 45 or 50 nt in length.

siRNA may have modifications internally, or at one or both ends. Examplemodifications at the 5′ end are illustrated in FIG. 1. These include:C6-alkyl (5′-hexylphosphate), 5′-Methyoxy; 5′-inverted dT (idT), and5′-beta-L-uridine. The modifications at the ends can help stabilize thesiRNA, protecting it from degradation by nucleases in the blood. ThesiRNAs may optionally be directed to regions of the HSF1 mRNA known orpredicted to be near or at splice sites of the gene; e.g., exon-intronjunctions. The siRNAs can also optionally be designed to anneal to knownor predicted exposed and/or single-stranded regions of the mRNA (e.g.,loops).

An antisense nucleic acid can be constructed using chemical synthesisand enzymatic ligation reactions using procedures known in the art. Forexample, an antisense nucleic acid (e.g., an antisense oligonucleotide)can be chemically synthesized using naturally-occurring nucleotides orvariously modified nucleotides designed to decrease off-target effects,and/or increase the biological stability of the molecules or to increasethe physical stability of the duplex formed between the antisense andsense nucleic acids. In at least one embodiment a modified sugarbackbone, including a phosphorothioate linkage or its derivatives, andacridine substituted nucleotides can be used.

Each of “G,” “C,” “A,” “T” and “U” generally stand for a nucleotide thatcontains guanine, cytosine, adenine, thymidine and uracil as a base,respectively. However, the term “ribonucleotide” or “nucleotide” canalso refer to a modified nucleotide or a surrogate replacement moiety.The skilled person is well aware that guanine, cytosine, adenine, anduracil may be replaced by other moieties without substantially alteringthe base pairing properties of an oligonucleotide comprising anucleotide bearing such replacement moiety. For example, withoutlimitation, a nucleotide comprising inosine as its base may base pairwith nucleotides containing adenine, cytosine, or uracil. Hence,nucleotides containing uracil, guanine, or adenine may be replaced inthe nucleotide sequences of dsRNA featured in the disclosure by anucleotide containing, for example, inosine. In another example, adenineand cytosine anywhere in the oligonucleotide can be replaced withguanine and uracil, respectively to form G-U Wobble base pairing withthe target mRNA. Sequences containing such replacement moieties aresuitable for the compositions and methods featured in the disclosure.

Modifications

The skilled artisan will recognize that the term “RNA molecule” or“ribonucleic acid molecule” encompasses not only RNA molecules asexpressed or found in nature, but also analogs and derivatives of RNAcomprising one or more ribonucleotide/ribonucleoside analogs orderivatives as described herein or as known in the art. Strictlyspeaking, a “ribonucleoside” includes a nucleoside base and a ribosesugar, and a “ribonucleotide” is a ribonucleoside with one, two or threephosphate moieties. However, the terms “ribonucleoside” and“ribonucleotide” can be considered to be equivalent as used herein. TheRNA can be modified in the nucleobase structure or in theribose-phosphate backbone structure, e.g., as described herein below.However, the molecules comprising ribonucleoside analogs or derivativesmust retain the ability to form a duplex. As non-limiting examples, anRNA molecule can also include at least one modified ribonucleoside,including but not limited to a 2′-O-methyl modified nucleotide, anucleoside comprising a 5′ phosphorothioate linkage group, a terminalnucleoside linked to a cholesteryl derivative or dodecanoic acidbisdecylamide group, a locked nucleoside, an abasic nucleoside, a2′-deoxy-2′-fluoro modified nucleoside, a 2′-amino-modified nucleoside,2′-alkyl-modified nucleoside, morpholino nucleoside, an unlockedribonucleotide (e.g., an acyclic nucleotide monomer, as described in WO2008/147824), a phosphoramidate or a non-natural base comprisingnucleoside, or any combination thereof Alternatively, an RNA moleculecan comprise at least two modified ribonucleosides, at least 3, at least4, at least 5, at least 6, at least 7, at least 8, at least 9, at least10, at least 15, at least 20 or more, up to the entire length of thedsRNA molecule. The modifications need not be the same for each of sucha plurality of modified ribonucleosides in an RNA molecule. In oneembodiment, modified RNAs contemplated for use in methods andcompositions described herein are peptide nucleic acids (PNAs) that havethe ability to form the required duplex structure and that permit ormediate the specific degradation of a target RNA via a RISC pathway.

Examples of modified nucleotides which can be used to generate theantisense nucleic acid include 5-fluorouracil, 5-bromouracil,5-chlorouracil, 5-iodouracil, hypoxanthine, xantine, 4-acetylcytosine,5-(carboxyhydroxylmethyl) uracil,5-carboxymethylaminomethyl-2-thiouridine,5-carboxymethylaminomethyluracil, dihydrouracil,beta-D-galactosylqueosine, inosine, N6-isopentenyladenine,1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine,2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine,7-methylguanine, 5-methylaminomethyluracil,5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine,5′-methoxycarboxymethyluracil, 5-methoxyuracil,2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v),wybutoxosine, pseudouracil, queosine, 2-thiocytosine,5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil,uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v),5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w,and 2,6-diaminopurine.

In one aspect, a modified ribonucleoside includes a deoxyribonucleoside.In such an instance, an iRNA agent can comprise one or moredeoxynucleosides, including, for example, a deoxynucleoside overhang(s),or one or more deoxynucleosides within the double stranded portion of adsRNA. However, it is self-evident that under no circumstances is adouble stranded DNA molecule encompassed by the term “iRNA.”

Replacing the 3′-terminal nucleotide overhanging segments of a 21-mersiRNA duplex having two-nucleotide 3′-overhangs withdeoxyribonucleotides does not have an adverse effect on RNAi activity.Replacing up to four nucleotides on each end of the siRNA withdeoxyribonucleotides has been well tolerated, whereas completesubstitution with deoxyribonucleotides results in no RNAi activity.International PCT Publication No. WO 00/44914, and Beach et al.International PCT Publication No. WO 01/68836 preliminarily suggest thatsiRNA may include modifications to either the phosphate-sugar backboneor the nucleoside to include at least one of a nitrogen or sulfurheteroatom. Kreutzer et al. Canadian Patent Application No. 2,359,180,also describe certain chemical modifications for use in dsRNA constructsin order to counteract activation of double-stranded RNA-dependentprotein kinase PKR, specifically 2′-amino or 2′-O-methyl nucleotides,and nucleotides containing a 2′-O or 4′-C methylene bridge. Additional3′-terminal nucleotide overhangs include dT (deoxythimidine),2′-O,4′-C-ethylene thymidine (eT), and 2-hydroxyethyl phosphate (hp).

Parrish et al. (2000 Molecular Cell 6: 1077-1087) tested certainchemical modifications targeting the unc-22 gene in C. elegans usinglong (>25 nt) siRNA transcripts. The authors describe the introductionof thiophosphate residues into these siRNA transcripts by incorporatingthiophosphate nucleotide analogs with T7 and T3 RNA polymerase andobserved that RNAs with two phosphorothioate modified bases also hadsubstantial decreases in effectiveness as RNAi. Further, Parrish et al.reported that phosphorothioate modification of more than two residuesgreatly destabilized the RNAs in vitro such that interference activitiescould not be assayed. Id. at 1081. The authors also tested certainmodifications at the 2′-position of the nucleotide sugar in the longsiRNA transcripts and found that substituting deoxynucleotides forribonucleotides produced a substantial decrease in interferenceactivity, especially in the case of Uridine to Thymidine and/or Cytidineto deoxy-Cytidine substitutions. Id. In addition, the authors testedcertain base modifications, including substituting, in sense andantisense strands of the siRNA, 4-thiouracil, 5-bromouracil,5-iodouracil, and 3-(aminoallyl)uracil for uracil, and inosine forguanosine. Whereas 4-thiouracil and 5-bromouracil substitution appearedto be tolerated, Parrish reported that inosine produced a substantialdecrease in interference activity when incorporated in either strand.Parrish also reported that incorporation of 5-iodouracil and3-(aminoallyl)uracil in the antisense strand resulted in a substantialdecrease in RNAi activity as well.

Those skilled in the art will appreciate that it is possible tosynthesize and modify the siRNA as desired, using any conventionalmethod known in the art (see Henschel et al. 2004 DEQOR: a web-basedtool for the design and quality control of siRNAs. Nucleic AcidsResearch 32 (Web Server Issue): W113-W120). Further, it will be apparentto those skilled in the art that there are a variety of regulatorysequences (for example, constitutive or inducible promoters,tissue-specific promoters or functional fragments thereof, etc.) whichare useful for the antisense oligonucleotide, siRNA, or shRNA expressionconstruct/vector.

There are several examples in the art describing sugar, base, phosphateand backbone modifications that can be introduced into nucleic acidmolecules with significant enhancement in their nuclease stability andefficacy. For example, oligonucleotides are modified to enhancestability and/or enhance biological activity by modification withnuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-flouro,2′-O-methyl, 2′-O-allyl, 2′-H, nucleotide base modifications (for areview see Usman and Cedergren 1992 TIBS. 17: 34; Usman et al. 1994Nucleic Acids Symp. Ser. 31: 163; Burgin et al. 1996 Biochemistry 35:14090). Sugar modifications of nucleic acids have been extensivelydescribed in the art.

Additional modifications and conjugations of RNAi agents have beendescribed. Soutschek et al. 2004 Nature 432: 173-178 presentedconjugation of cholesterol to the 3′-end of the sense strand of a siRNAmolecule by means of a pyrrolidine linker, thereby generating a covalentand irreversible conjugate. Chemical modifications (includingconjugation with other molecules) of siRNA may also be made to improvethe in vivo pharmacokinetic retention time and efficiency.

In various embodiments, the RNAi agent to HSF1 comprises at least one5′-uridine-adenine-3′ (5′-ua-3′) dinucleotide, wherein the uridine is a2′-modified nucleotide; at least one 5′-uridine-guanine-3′ (5′-ug-3′)dinucleotide, wherein the 5′-uridine is a 2′-modified nucleotide; leastone 5′-cytidine-adenine-3′ (5′-ca-3′) dinucleotide, wherein the5′-cytidine is a 2′-modified nucleotide; or at least one5′-uridine-uridine-3′ (5′-uu-3′) dinucleotide, wherein the 5′-uridine isa 2′-modified nucleotide.

In various embodiments, the RNAi agent comprises a 2′-modificationselected from the group consisting of: 2′-deoxy, 2′-deoxy-2′-fluoro,2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP),2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl(2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), and2′-O—N-methylacetamido (2′-O—NMA).

In another embodiment, the RNAi comprises a gap or missing base. Forexample, the phosphate-sugar backbone may be present, but the basemissing.

In another embodiment, the RNAi agent has a single-stranded nick (e.g.,a break or missing bond in the backbone). This nick can be, for example,in the sense strand, producing a small internally segmented interferingRNA, or sisiRNA, which may have less off-target effects than thecorresponding RNAi agent without a nick.

The antisense nucleic acid or RNAi agent can also have an alternativebackbone such as locked nucleic acids (LNA), Morpholinos, peptidicnucleic acids (PNA), threose nucleic acid (TNA), or glycol nucleic acid(GNA), and/or it can be labeled (e.g., radiolabeled or otherwisetagged). One or both strands can comprise an alternative backbone

In yet another embodiment, the antisense nucleic acid molecule employedby the methods of the present disclosure can include an a-anomericnucleic acid molecule. An α-anomeric nucleic acid molecule formsspecific double-stranded hybrids with complementary RNA in which,contrary to the usual β-units, the strands run parallel to each other.Gaultier et al. 1987 Nucleic Acids. Res. 15: 6625-6641. The antisensenucleic acid molecule can also comprise a 2′-o-methylribonucleotide(Inoue et al. 1987 Nucleic Acids Res. 15: 6131-6148) or a chimericRNA-DNA analogue (Inoue et al. 1987 FEBS Lett. 215: 327-330).

In still another embodiment, an antisense nucleic acid is a ribozyme.Ribozymes are catalytic RNA molecules with ribonuclease activity whichare capable of cleaving a single-stranded nucleic acid, such as an mRNA,to which they have a complementary region. Thus, ribozymes [e.g.,hammerhead ribozymes (described in Haselhoff et al. 1988, Nature 334:585-591)] can be used to catalytically cleave HSF1 mRNA transcripts tothereby inhibit translation of HSF1 mRNA.

Alternatively, gene expression can be inhibited by targeting nucleotidesequences complementary to the regulatory region of HSF1 (e.g., thepromoter and/or enhancers) to form triple helical structures thatprevent transcription of the HSF1 gene. See generally, Helene 1991Anticancer Drug Des. 6(6): 569-84; Helene et al. 1992 Ann. N.Y. Acad.Sci. 660: 27-36; and Maher 1992, Bioassays 14(12): 807-15.

Alternatively, the antisense nucleic acid can be produced biologicallyusing an expression vector into which a nucleic acid has been subclonedin an antisense orientation (i.e., RNA transcribed from the insertednucleic acid will be in an antisense orientation to a target nucleicacid of interest).

The antisense nucleic acid molecules of the present disclosure aretypically administered to a subject or generated in situ such that theyhybridize with cellular mRNA and/or genomic DNA encoding HSF1, andinhibit expression by inhibiting transcription and/or translation. Anexample of a route of administration of antisense nucleic acid moleculesincludes direct injection at a tissue site. Alternatively, antisensenucleic acid molecules can be modified to target selected cells and thenadministered systemically. For example, for systemic administration,antisense molecules can be modified such that they specifically bind toreceptors or antigens expressed on a selected cell surface, e.g., bylinking the antisense nucleic acid molecules to peptides or antibodieswhich bind to cell surface receptors or antigens. The antisense nucleicacid molecules can also be delivered to cells using vectors well knownin the art and described in, for example, US20070111230, the entirecontents of which are incorporated herein. To achieve sufficientintracellular concentrations of the antisense molecules, vectorconstructs in which the antisense nucleic acid molecule is placed underthe control of a strong pol II or pol III promoter can be used.

RNA Interference

RNAi (RNA interference) has been studied in a variety of systems. Recentwork in Drosophila embryonic lysates (Elbashir et al. 2001 EMBO J. 20:6877 and Tuschl et al. International PCT Publication No. WO 01/75164)has revealed certain requirements for siRNA length, structure, chemicalcomposition, and sequence that are essential to mediate efficient RNAiactivity. These studies have shown that 21-nucleotide siRNA duplexes aremost active when containing 3′-terminal dinucleotide overhangs.Substitution of the 3′-terminal siRNA overhang nucleotides with 2′-deoxynucleotides (2′-H) was tolerated. In addition, a 5′-phosphate on thetarget-complementary strand of an siRNA duplex is usually required forsiRNA activity.

The use of longer dsRNA has been described. For example, Beach et al.International PCT Publication No. WO 01/68836, describes attenuatinggene expression using endogenously-derived dsRNA. Tuschl et al.International PCT Publication No. WO 01/75164, describe a Drosophila invitro RNAi system and the use of specific siRNA molecules for certainfunctional genomic and certain therapeutic applications. Li et al.International PCT Publication No. WO 00/44914, describe the use ofspecific long (141 bp-488 bp) enzymatically synthesized or vectorexpressed dsRNAs for attenuating the expression of certain target genes.Zernicka-Goetz et al. International PCT Publication No. WO 01/36646,describe certain methods for inhibiting the expression of particulargenes in mammalian cells using certain long (550 bp-714 bp),enzymatically synthesized or vector expressed dsRNA molecules. Fire etal. International PCT Publication No. WO 99/32619, describe particularmethods for introducing certain long dsRNA molecules into cells for usein inhibiting gene expression in nematodes. Plaetinck et al.International PCT Publication No. WO 00/01846, describe certain methodsfor identifying specific genes responsible for conferring a particularphenotype in a cell using specific long dsRNA molecules. Mello et al.International PCT Publication No. WO 01/29058, describe theidentification of specific genes involved in dsRNA-mediated RNAi.Pachuck et al. International PCT Publication No. WO 00/63364, describecertain long (at least 200 nt) dsRNA constructs. Deschamps Depailletteet al. International PCT Publication No. WO 99/07409, describe specificcompositions consisting of particular dsRNA molecules combined withcertain anti-viral agents. Waterhouse et al. International PCTPublication No. 99/53050 and 1998, PNAS, 95, 13959-13964, describecertain methods for decreasing the phenotypic expression of a nucleicacid in plant cells using certain dsRNAs. Driscoll et al. InternationalPCT Publication No. WO 01/49844, describe specific DNA expressionconstructs for use in facilitating gene silencing in targeted organisms.

Others have reported on various RNAi and gene-silencing systems. Forexample, Parrish et al. 2000, Molecular Cell 6: 1077-1087 describesspecific chemically modified dsRNA constructs targeting the unc-22 geneof C. elegans. Grossniklaus, International PCT Publication No. WO01/38551, describes certain methods for regulating polycomb geneexpression in plants using certain dsRNAs. Churikov et al. InternationalPCT Publication No. WO 01/42443, describe certain methods for modifyinggenetic characteristics of an organism using certain dsRNAs. Cogoni etal, International PCT Publication No. WO 01/53475, describe certainmethods for isolating a Neurospora silencing gene and uses thereof Reedet al. International PCT Publication No. WO 01/68836, describe certainmethods for gene silencing in plants. Honer et al.

International PCT Publication No. WO 01/70944, describe certain methodsof drug screening using transgenic nematodes as Parkinson's Diseasemodels using certain dsRNAs. Deak et al. International PCT PublicationNo. WO 01/72774, describe certain Drosophila-derived gene products thatmay be related to RNAi in Drosophila. Arndt et al. International PCTPublication No. WO 01/92513 describe certain methods for mediating genesuppression by using factors that enhance RNAi. Tuschl et al.International PCT Publication No. WO 02/44321, describe certainsynthetic siRNA constructs. Pachuk et al. International PCT PublicationNo. WO 00/63364, and Satishchandran et al. International PCT PublicationNo. WO 01/04313, describe certain methods and compositions forinhibiting the function of certain polynucleotide sequences usingcertain long (over 250 bp), vector expressed dsRNAs. Echeverri et al.International PCT Publication No. WO 02/38805, describe certain C.elegans genes identified via RNAi. Kreutzer et al. International PCTPublications Nos. WO 02/055692, WO 02/055693, and EP 1144623 B1describes certain methods for inhibiting gene expression using dsRNA.Graham et al. International PCT Publications Nos. WO 99/49029 and WO01/70949, and AU 4037501 describe certain vector expressed siRNAmolecules. Fire et al. U.S. Pat. No. 6,506,559, describe certain methodsfor inhibiting gene expression in vitro using certain long dsRNA (299bp-1033 bp) constructs that mediate RNAi. Martinez et al. 2002, Cell,110, 563-574, describe certain single-stranded siRNA constructs,including certain 5′-phosphorylated single-stranded siRNAs that mediateRNA interference in HeLa cells. Harborth et al. 2003, Antisense &Nucleic Acid Drug Development, 13, 83-105, describe certain chemicallyand structurally modified siRNA molecules. Chiu and Rana, 2003, RNA, 9,1034-1048, describe certain chemically and structurally modified siRNAmolecules. Woolf et al. International PCT Publication Nos. WO 03/064626and WO 03/064625 describe certain chemically modified dsRNA constructs.

In various embodiments, the RNAi agent to HSF1 is ligated to one or morediagnostic compound, reporter group, cross-linking agent,nuclease-resistance conferring moiety, natural or unusual nucleobase,lipophilic molecule, cholesterol, lipid, lectin, steroid, uvaol,hecigenin, diosgenin, terpene, triterpene, sarsasapogenin, Friedelin,epifriedelanol-derivatized lithocholic acid, vitamin, carbohydrate,dextran, pullulan, chitin, chitosan, synthetic carbohydrate, oligolactate 15-mer, natural polymer, low- or medium-molecular weightpolymer, inulin, cyclodextrin, hyaluronic acid, protein, protein-bindingagent, integrin-targeting molecule, polycationic, peptide, polyamine,peptide mimic, and/or transferrin.

Delivery of RNAi Agents

RNAi agents of the present disclosure can be delivered or introduced(e.g., to a cell in vitro or to a patient) by any means known in theart.

“Introducing into a cell,” when referring to an iRNA, means facilitatingor effecting uptake or absorption into the cell, as is understood bythose skilled in the art. Absorption or uptake of an iRNA can occurthrough unaided diffusive or active cellular processes, or by auxiliaryagents or devices. The meaning of this term is not limited to cells invitro; an iRNA may also be “introduced into a cell,” wherein the cell ispart of a living organism. In such an instance, introduction into thecell will include the delivery to the organism. For example, for in vivodelivery, iRNA can be injected into a tissue site or administeredsystemically. In vivo delivery can also be by a beta-glucan deliverysystem, such as those described in U.S. Pat. Nos. 5,032,401 and5,607,677, and U.S. Publication No. 2005/0281781 which are herebyincorporated by reference in their entirety. In vitro introduction intoa cell includes methods known in the art such as electroporation andlipofection. Further approaches are described below or known in the art.

Delivery of RNAi agent to tissue is a problem both because the materialmust reach the target organ and must also enter the cytoplasm of targetcells. RNA cannot penetrate cellular membranes, so systemic delivery ofnaked RNAi agent is unlikely to be successful. RNA is quickly degradedby RNAse activity in serum. For these reasons, other mechanisms todeliver RNAi agent to target cells has been devised. Methods known inthe art include but are not limited to: viral delivery (retrovirus,adenovirus, lentivirus, baculovirus, AAV); liposomes (Lipofectamine,cationic DOTAP, neutral DOPC) or nanoparticles (cationic polymer, PEI),bacterial delivery (tkRNAi), and also chemical modification (LNA) ofsiRNA to improve stability. Xia et al. 2002 Nat. Biotechnol. 20 andDevroe et al. 2002. BMC Biotechnol. 21: 15, disclose incorporation ofsiRNA into a viral vector. Other systems for delivery of RNAi agents arecontemplated, and the RNAi agents of the present disclosure can bedelivered by various methods yet to be found and/or approved by the FDAor other regulatory authorities.

Liposomes have been used previously for drug delivery (e.g., delivery ofa chemotherapeutic). Liposomes (e.g., cationic liposomes) are describedin PCT publications WO02/100435A1, WO03/015757A1, and WO04029213A2; U.S.Pat. Nos. 5,962,016; 5,030,453; and 6,680,068; and U.S. PatentApplication 2004/0208921. A process of making liposomes is alsodescribed in WO04/002453A1. Furthermore, neutral lipids have beenincorporated into cationic liposomes (e.g., Farhood et al. 1995).Cationic liposomes have been used to deliver RNAi agent to various celltypes (Sioud and Sorensen 2003; U.S. Patent Application 2004/0204377;Duxbury et al., 2004; Donze and Picard, 2002). Use of neutral liposomesdisclosed in Miller et al. 1998, and U.S. Publ. 2003/0012812.

As used herein, the term “SNALP” refers to a stable nucleic acid-lipidparticle. A

SNALP represents a vesicle of lipids coating a reduced aqueous interiorcomprising a nucleic acid such as an iRNA or a plasmid from which aniRNA is transcribed. SNALPs are described, e.g., in U.S. PatentApplication Publication Nos. 20060240093, 20070135372, and inInternational Application No. WO 2009082817. These applications areincorporated herein by reference in their entirety.

Chemical transfection using lipid-based, amine-based and polymer-basedtechniques, is disclosed in products from Ambion Inc., Austin, Tex.; andNovagen, EMD Biosciences, Inc, an Affiliate of Merck KGaA, Darmstadt,Germany); Ovcharenko D (2003) “Efficient delivery of siRNAs to humanprimary cells.” Ambion TechNotes 10 (5): 15-16). Additionally, Song etal. (Nat Med. published online (Fete 10, 2003) doi: 10.1038/nm828) andothers [Caplen et al. 2001 Proc. Natl. Acad. Sci. (USA), 98: 9742-9747;and McCaffrey et al. Nature 414: 34-39] disclose that liver cells can beefficiently transfected by injection of the siRNA into a mammal'scirculatory system.

A variety of molecules have been used for cell-specific RNAi agentdelivery. For example, the nucleic acid-condensing property of protaminehas been combined with specific antibodies to deliver siRNAs. Song etal. 2005 Nat Biotch. 23: 709-717. The self-assembly PEGylated polycationpolyethylenimine (PEI) has also been used to condense and protectsiRNAs. Schiffelers et al. 2004 Nucl. Acids Res. 32: e149, 141-110.

The siRNA-containing nanoparticles were then successfully delivered tointegrin-overexpressing tumor neovasculature. Hu-Lieskovan et al. 2005Cancer Res. 65: 8984-8992.

The RNAi agents of the present disclosure can be delivered via, forexample, Lipid nanoparticles (LNP); neutral liposomes (NL); polymernanoparticles; double-stranded RNA binding motifs (dsRBMs); or viamodification of the RNAi agent (e.g., covalent attachment to the dsRNA).

Lipid nanoparticles (LNP) are self-assembling cationic lipid basedsystems. These can comprise, for example, a neutral lipid (the liposomebase); a cationic lipid (for siRNA loading); cholesterol (forstabilizing the liposomes); and PEG-lipid (for stabilizing theformulation, charge shielding and extended circulation in thebloodstream). The cationic lipid can comprise, for example, a headgroup,a linker, a tail and a cholesterol tail. The LNP can have, for example,good tumor delivery, extended circulation in the blood, small particles(e.g., less than 100 nm), and stability in the tumor microenvironment(which has low pH and is hypoxic).

Neutral liposomes (NL) are non-cationic lipid based particles.

Polymer nanoparticles are self-assembling polymer-based particles.

Double-stranded RNA binding motifs (dsRBMs) are self-assembling RNAbinding proteins, which will need modifications.

RNAi Agents to HSF1

siRNAs that are particularly useful for this disclosure include thosewhich can bind specifically to a region of the HSF1 mRNA, and have oneor more of the following qualities: binding in the coding segment ofHSF1; binding at or near the junction of the 5′ untranslated region andthe start of the coding segment; binding at or near the translationalstart site of the mRNA; binding at or near junctions of exons andintrons; little or no binding to the mRNAs of other genes (little or no“off-target effects”); binding to the HSF1 mRNA in or near a region orregions that is not double-stranded or a stem region, e.g., in a loop orsingle-stranded portion; eliciting little or no immunogenicity; bindingin a segment of the HSF1 mRNA sequence which is conserved among variousanimal species (including human, mouse, rat, cynomolgus monkey, etc.),as the presence of a conserved sequence facilitates testing usingvarious laboratory animals; binding to double-stranded region(s) of themRNA; binding to an AT-rich region (e.g., at least about 50, 51, 52, 53,54, 55, 56, 57, 58, 59, or 60% AT-rich); and lacking particularsequences known or suspected to decrease siRNA activity, e.g., thepresence of a GG sequence at the 5′ end, which may decrease separationof the double-stranded portion of the siRNA.

RNAi agents can be designed as HSF1 RNAi agents which bind to and assistin degradation of HSF1 mRNA. The anti-HSF1 RNAi agents can be designedto bind to the coding segment or non-coding segment (e.g., the 5′ or 3′untranslated regions, or UTRs). Preferably the RNAi agent binds to thecoding segment of the mRNA. The RNAi agents can have double-strandedregions of, for example, about 17, 18, 19, 20, 21, 22, 23, or 24 bp.Preferably the RNAi agent comprises about 19, 20 or 21 bp. The RNAiagents can be longer (e.g., up to 49 bp), as incorporated into aconstruct suitable for shortening by the Dicer complex. The RNAi canalso be incorporated into a longer construct for expression prior tofurther shortening and processing.

Sense and Antisense Strand of HSF1 RNAi Agents

The RNAi agents comprise a first strand and a second strand. In oneembodiment, the first and second strands are a sense strand and anantisense strand, respectively. In other embodiments, the first andsecond strands are an antisense and sense strand, respectively. Thefirst strand thus can comprise a sense or antisense strand of anysequence disclosed herein, or a variant sequence thereof comprising 15contiguous nucleotides with up to 3 mismatches; and the second strandcan thus comprise the corresponding antisense or sense strand of anysequence disclosed herein, or a variant sequence thereof comprising 15contiguous nucleotides with up to 3 mismatches.

The term “antisense strand” refers to the strand of an iRNA, e.g., adsRNA, which includes a region that is substantially complementary to atarget sequence. As used herein, the term “region of complementarity”refers to the region on the antisense strand that is substantiallycomplementary to a sequence, for example a target sequence, as definedherein. Where the region of complementarity is not fully complementaryto the target sequence, the mismatches may be in the internal orterminal regions of the molecule. Generally, the most toleratedmismatches are in the terminal regions, e.g., within 5, 4, 3, or 2nucleotides of the 5′ and/or 3′ terminus

The term “sense strand,” as used herein, refers to the strand of an iRNAthat includes a region that is substantially complementary to a regionof the antisense strand as that term is defined herein.

Overhangs and Blunt Ends

The RNAi agents can have 0, 1, or 2 overhangs; in the case of 0 ntoverhangs, both ends are blunt-ended. An RNAi agent can have 0, 1 or 2blunt ends. In a “blunt-ended RNAi agent” no strands contain unpairednucleotides at that end; thus a blunt-ended molecule lacks either 3′ or5′ single-stranded nucleotide overhangs.

As used herein, the term “overhang” or “nucleotide overhang” refer to atleast one unpaired nucleotide that protrudes from the duplex structureof an iRNA, e.g., a dsRNA. For example, when a 3′-end of one strand of adsRNA extends beyond the 5′-end of the other strand, or vice versa,there is an overhang. A dsRNA can comprise an overhang of at least onenucleotide; alternatively the overhang can comprise at least 2 nt, atleast 3 nt, at least 4 nt, at least 5 nt or more. An overhang cancomprise or consist of a nucleotide/nucleoside analog, including adeoxynucleotide/nucleoside. The overhang(s) may be on the sense strand,the antisense strand or any combination thereof The nucleotide(s) of anoverhang can be present on the 5′ end, 3′ end or both ends of either anantisense or sense strand of a dsRNA.

The terms “blunt” or “blunt-ended” as used herein in reference to adsRNA mean that there are no unpaired nucleotides or nucleotide analogsat a given terminal end of a dsRNA, i.e., no nucleotide overhang. One orboth ends of a dsRNA can be blunt. Where both ends of a dsRNA are blunt,the dsRNA is said to be blunt ended. To be clear, a “blunt ended” dsRNAis a dsRNA that is blunt at both ends, i.e., no nucleotide overhang ateither end of the molecule. Most often such a molecule will bedouble-stranded over its entire length.

In one embodiment, a blunt end of an RNAi duplex is chemically modifiedby the addition of a 3′ cap, e.g., those described in WO 2005/021749 andWO 2007/128477. In such embodiments, the 3′ caps are non-nucleotidic,and thus do not constitute an overhang.

The mRNA sequence of a gene may vary from individual to individual,especially at wobble positions within the coding segment, or in theuntranslated region; individuals may also differ from each other incoding sequence, resulting in additional differences in mRNA andcorresponding RNAi agent sequence. RNAi agents can also be modified insequence to reduce immunogenicity, binding to undesired genes (e.g.,“off-target effects”) or to increase stability in the blood. (Thesesequence variants are independent of chemical modification of the basesor 5′ or 3′ or other end-caps of the RNAi agents.)

Example anti-HSF1 RNAi agents include those which bind to an HSF1 geneprovided herein. Example siRNAs to HSF1 are provided in any one or moreof Tables 1, 2, 3, 3A, 8, 9A and 9B.

Measuring the Effect of an RNAi Agent on HSF1 Activity, Level and/orExpression

Any method known in the art can be use to measure changes in HSF1activity, level and/or expression induced by a HSF1 siRNA. Measurementscan be performed at multiple timepoints, prior to, during and afteradministration of the siRNA, to determine the effect of the siRNA.

The RNAi agents of the present disclosure silence, inhibit theexpression of, down-regulate the expression of, and/or suppress theexpression of HSF1.

The terms “silence,” “inhibit the expression of,” “down-regulate theexpression of,” “suppress the expression of,” and the like, in so far asthey refer to a HSF1 gene, herein refer to the at least partialsuppression of the expression of a HSF1 gene, as manifested by areduction of the amount of HSF1 mRNA which may be isolated from ordetected in a first cell or group of cells in which a HSF1 gene istranscribed and which has or have been treated such that the expressionof a HSF1 gene is inhibited, as compared to a second cell or group ofcells substantially identical to the first cell or group of cells butwhich has or have not been so treated (control cells). The degree ofinhibition is usually expressed in terms of

$\begin{matrix}{{\frac{\left( {{mRNA}\mspace{14mu} {in}\mspace{14mu} {control}\mspace{14mu} {cells}} \right) - \left( {{mRNA}\mspace{14mu} {in}\mspace{14mu} {treated}\mspace{14mu} {cells}} \right)}{\left( {{mRNA}\mspace{14mu} {in}\mspace{14mu} {control}\mspace{14mu} {cells}} \right)} \cdot 100}\%} & \left( {{Equation}\mspace{14mu} 1} \right)\end{matrix}$

Alternatively, the degree of inhibition may be given in terms of areduction of a parameter that is functionally linked to HSF1 geneexpression, e.g., the amount of protein encoded by a HSF1 gene, or thenumber of cells displaying a certain phenotype, e.g., modulation ofexpression of a gene (e.g., HSP70) whose expression is mediated in wholeor in part by HSF1. In principle, HSF1 gene silencing may be determinedin any cell expressing HSF1, either constitutively or by genomicengineering, and by any appropriate assay. However, when a reference orcontrol is needed in order to determine whether a given iRNA inhibitsthe expression of HSF1 by a certain degree and therefore is encompassedby the instant disclosure, the assays provided in the Examples belowshall serve as such reference.

For example, in certain instances, expression of a HSF1 gene issuppressed by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or50% by administration of an iRNA featured in the disclosure. In someembodiments, a HSF1 gene is suppressed by at least about 60%, 70%, or80% by administration of an iRNA featured in the disclosure. In someembodiments, a HSF1 gene is suppressed by at least about 85%, 90%, or95% or more by administration of an iRNA as described herein.

Treatments Involving HSF1 RNAi Agents

As used herein in the context of HSF1 expression, the terms “treat,”“treatment,” and the like, refer to relief from or alleviation ofpathological processes mediated by HSF1 expression. In the context ofthe present disclosure insofar as it relates to any of the otherconditions recited herein below (other than pathological processesmediated by HSF1 expression), the terms “treat,” “treatment,” and thelike mean to relieve or alleviate at least one symptom associated withsuch condition, or to slow or reverse the progression or anticipatedprogression of such condition, such as slowing the progression of alipid disorder, such as atherosclerosis.

By “lower” in the context of a disease marker or symptom is meant astatistically significant decrease in such level. The decrease can be,for example, at least 10%, at least 20%, at least 30%, at least 40% ormore. If, for a particular disease, or for an individual suffering froma particular disease, the levels or expression of HSF1 are elevated,treatment with an HSF1 RNAi agent of the present disclosure canpreferably reduce the level or expression of HSF1 to a level consideredin the literature as within the range of normal for an individualwithout such disorder.

The level or expression of HSF1 can be measured by evaluation of mRNA(e.g., via Northern blots or PCR), or protein (e.g., Western blots). Theeffect of an RNAi agent on HSF1 expression can be determined bymeasuring HSF1 gene transcription rates (e.g., via Northern blots; orreverse transcriptase polymerase chain reaction or real-time polymerasechain reaction). RT-PCR has been used to show that mRNA levels of HSF1are high in kidney, pancreas and prostate, and medium in liver andspleen. Brauner-Osborne et al. 2001. Biochim. Biophys. Acta 1518:237-248. Direct measurements can be made of levels of HSF1 (which isexpressed by the cell surface), e.g. by Western blots of tissues inwhich HSF1 is expressed.

In another embodiment of the disclosure, the compositions comprising aHSF1 RNAi agent can be administered to non-human animals. For example,the compositions can be given to chickens, turkeys, livestock animals(such as sheep, pigs, horses, cattle, etc.), companion animals (e.g.,cats and dogs) and can have efficacy in treatment of cancer and viraldiseases. In each case, the RNAi agent to HSF1 would be selected tomatch the sequence of the HSF1 of the genome of the animal, and to,preferably, contain at least 1 nt mismatch from all other genes in thatanimal's genome.

By “treatment” is meant prophylaxis, therapy, cure, or any other changein a patient's condition indicating improvement or absence ofdegradation of physical condition. By “treatment” is meant treatment ofHSF1-related disease (e.g., cancer or viral disease), or any appropriatetreatment of any other ailment the patient has. As used herein, theterms “treatment” and “treat” refer to both prophylactic or preventativetreatment and curative or disease-modifying treatment, includingtreatment of patients at risk of contracting a disease or suspected ofhaving a disease, as well as patients already ill or diagnosed assuffering from a condition. The terms “treatment” and “treat” also referto the maintenance and/or promotion of health in an individual notsuffering from a disease but who may be susceptible to developing anunhealthy condition, such as nitrogen imbalance or muscle loss. In oneembodiment, “treatment” does not encompass prevention of a diseasestate. Thus, the present disclosure is useful for suppressing expressionof HSF1 and/or treating an HSF1-related disease in an individualafflicted by an HSF1-related disease, or an individual susceptible to anHSF1-related disease. An individual “afflicted” by an HSF1-relateddisease has demonstrated detectable symptoms characteristics of thedisease, or had otherwise been shown clinically to have been exposed toor to carry HSF1-related disease pathogens or markers. As non-limitingexamples, an individual afflicted by an HSF1-related disease can showoutward symptoms; or can show no outward symptoms but can be shown witha clinical test to carry protein markers associated with an HSF1-relateddisease, or proteins or genetic material associated with a pathogen inthe blood.

An “effective amount” or a “therapeutically effective amount” is anamount that treats a disease or medical condition of an individual, or,more generally, provides a nutritional, physiological or medical benefitto an individual. As used herein, the phrases “therapeutically effectiveamount” and “prophylactically effective amount” refer to an amount thatprovides a therapeutic benefit in the treatment, prevention, ormanagement of pathological processes mediated by HSF1 expression or anovert symptom of pathological processes mediated by HSF1 expression. Thespecific amount that is therapeutically effective can be readilydetermined by an ordinary medical practitioner, and may vary dependingon factors known in the art, such as, e.g., the type of pathologicalprocesses mediated by HSF1 expression, the patient's history and age,the stage of pathological processes mediated by HSF1 expression, andadministration of other agents that inhibit pathological processesmediated by HSF1.

In various embodiments of the disclosure, the patient is at least about1, 5, 10, 20, 30, 40, 50, 55, 60, 65, 70, or 75 years of age. In variousembodiments, the patient is no more than about 1, 5, 10, 20, 30, 40, 50,55, 60, 65, 70, 75, 80, 90, or 100 years of age. In various embodimentsthe patient has a body weight of at least about 20, 30, 40, 50, 60, 70,80, 90, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340,360, 380 or 400 lbs. In various embodiments, the patient has a bodyweight of no more than about 20, 30, 40, 50, 60, 70, 80, 90, 100, 120,140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380 or 400lbs.

In various embodiments of the disclosure, the dosage [measuring only theactive ingredient(s)] can be at least about 1, 5, 10, 25, 50, 100, 200,250, 300, 250, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900,950 or 1000 ng, 1, 5, 10, 25, 50, 100, 200, 250, 300, 250, 400, 450,500, 550, 600, 650, 700, 750, 800, 850, 900, 950 or 1000 micrograms, 1,5, 10, 25, 50, 100, 200, 250, 300, 250, 400, 450, 500, 550, 600, 650,700, 750, 800, 850, 900, 950 or 1000 mg. In various embodiments, thedosage can be no more than about 10, 25, 50, 100, 200, 250, 300, 250,400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950 or 1000 mg.In various embodiments, the dosage can be administered at least morethan once a day, daily, more than once a weekly, weekly, bi-weekly,monthly, and/or every 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or acombination thereof

In various embodiments, the dosage is correlated to the body weight orbody surface area of the individual. The actual dosage level can bevaried to obtain an amount of active agent which is effective for aparticular patient, composition and mode of administration, withoutbeing toxic to the patient. The selected dose will depend on a varietyof pharmacokinetic factors, including the activity of the particularRNAi agent employed, the route of administration, the rate of excretionof the RNAi agent, the duration of the treatment, other drugs, compoundsand/or materials used in combination with the RNAi agent, the age, sex,weight, condition, general health and prior medical history of thepatient, and like factors well known in the medical arts. A physician orveterinarian having ordinary skill in the art can readily determine theeffective amount of the RNAi agent required. A suitable dose will bethat amount which is the lowest dose effective to produce a therapeuticeffect, or a dose low enough to produce a therapeutic effect withoutcausing side effects.

Pharmaceutical Compositions Comprising a HSF1 RNAi Agent

As used herein, a “pharmaceutical composition” comprises apharmacologically effective amount of an iRNA and a pharmaceuticallyacceptable carrier. As used herein, “pharmacologically effectiveamount,” “therapeutically effective amount” or simply “effective amount”refers to that amount of an iRNA effective to produce the intendedpharmacological, therapeutic or preventive result. For example, if agiven clinical treatment is considered effective when there is at leasta 10% reduction in a measurable parameter associated with a disease ordisorder, a therapeutically effective amount of a drug for the treatmentof that disease or disorder is the amount necessary to effect at least a10% reduction in that parameter. For example, a therapeuticallyeffective amount of an iRNA targeting HSF1 can reduce HSF1 proteinlevels by at least 10%.

The term “pharmaceutically acceptable carrier” refers to a carrier foradministration of a therapeutic agent. Such carriers include, but arenot limited to, saline, buffered saline, dextrose, water, glycerol,ethanol, and combinations thereof. The term specifically excludes cellculture medium. For drugs administered orally, pharmaceuticallyacceptable carriers include, but are not limited to pharmaceuticallyacceptable excipients such as inert diluents, disintegrating agents,binding agents, lubricating agents, sweetening agents, flavoring agents,coloring agents and preservatives. Suitable inert diluents includesodium and calcium carbonate, sodium and calcium phosphate, and lactose,while corn starch and alginic acid are suitable disintegrating agents.Binding agents may include starch and gelatin, while the lubricatingagent, if present, will generally be magnesium stearate, stearic acid ortalc. If desired, the tablets may be coated with a material such asglyceryl monostearate or glyceryl distearate, to delay absorption in thegastrointestinal tract. Agents included in drug formulations aredescribed herein.

The pharmaceutical compositions comprising a HSF1 RNAi agent can be insolid form, for example, powders, granules, tablets, pills, gelcaps,gelatin capsules, liposomes, suppositories, chewable forms, or patches.The pharmaceutical compositions comprising a HSF1 RNAi agent can also bepresented in liquid form, for example, solutions, emulsions,suspensions, elixirs, or syrups. Appropriate liquid supports can be, forexample, water, organic solvents such as polyol, such as glycerol orglycols, including propylene glycol and polyethylene glycol, or ethanol,Cremophor EL, or mixtures thereof, in varying proportions, in water. Thecompositions can comprise nano-sized amorphous or crystalline granulescoated with albumin or a surfactant.

Appropriate supports can include, for example, antibacterial andantifungal agents, buffering agents, calcium phosphate, cellulose,methyl cellulose, chlorobutanol, cocoa butter, colorings, dextrin,emulsifiers, enteric coatings, flavorings, gelatin, isotonic agents,lecithin, magnesium stearate, perfuming agents, polyalcohols such asmannitol, injectable organic esters such as ethyl oleate, paraben,phenol sorbic acid, polyethylene glycol, polyvinylpyrrolidine, phosphatebuffered saline (PBS), preserving agents, propylene glycol, sodiumcarboxymethylcellulose, sodium chloride, sorbitol, various sugars(including, but not limited to, sucrose, fructose, galactose, lactoseand trehalose), starch, suppository wax, talc, vegetable oils, such asolive oil and corn oil, vitamins, wax, and/or wetting agents. For HSF1RNAi agents, a particular support comprises dextran and water, e.g. 5%dextrose in water (D5W).

The biologically inert portion of the pharmaceutical composition canoptionally be erodible, allowing timed release of the RNAi agent.

The pharmaceutical composition comprising a HSF1 can be administered bybuccal, inhalation (including insufflation and deep inhalation), nasal,oral, parenteral, implant, injection or infusion via epidural,intra-arterial, intra-articular, intracapsular, intracardiac,intracerebroventricular, intracranial, intradermal, intramuscular,intraorbital, intraperitoneal, intraspinal, intrasternal, intrathecal,intravenous, subarachnoid, subcapsular, subcutaneous, subcuticular,transendothelial, transtracheal, transvascular, rectal, sublingual,topical, and/or vaginal routes. This may be by injection, infusion,dermal patch, or any other method known in the art. The formulation canbe powdered, nebulized, aerosolized, granulized or otherwiseappropriately prepared for delivery. The administration, if liquid, maybe slow or via bolus, though, under some circumstances known in the art,bolus injections may lead to loss of material through the kidneys.

The HSF1 RNAi agents can be administered with medical devices known inthe art. For example, in a particular specific embodiment, an RNAi agentcan be administered with a needleless hypodermic injection device, suchas the devices disclosed in U.S. Pat. Nos. 5,399,163, 5,383,851,5,312,335, 5,064,413, 4,941,880, 4,790,824, or 4,596,556. Examples ofwell-known implants and modules useful in the present disclosureinclude: U.S. Pat. No. 4,487,603, which discloses an implantablemicro-infusion pump for dispensing medication at a controlled rate; U.S.Pat. No. 4.,486,194, which discloses a therapeutic device foradministering medications through the skin; U.S. Pat. No. 4,447,233,which discloses a medication infusion pump for delivering medication ata precise infusion rate; U.S. Pat. No. 4,447,224, which discloses avariable flow implantable infusion apparatus for continuous drugdelivery; U.S. Pat. No. 4,439,196, which discloses an osmotic drugdelivery system having multi-chamber compartments; and U.S. Pat. No.4,475,196, which discloses an osmotic drug delivery system. Many othersuch implants, delivery systems, and modules are known to those skilledin the art.

In certain embodiments, RNAi agents can be formulated to ensure properdistribution in vivo. For example, the blood-brain barrier (BBB)excludes many highly hydrophilic compounds. To ensure that the HSF1 RNAiagents cross the BBB (if desired), they can be formulated, for example,in liposomes. For methods of manufacturing liposomes, see, e.g., U.S.Pat. Nos. 4,522,811; 5,374,548; and 5,399,331. The liposomes maycomprise one or more moieties which are selectively transported intospecific cells or organs, thus enhance targeted drug delivery (see,e.g., V. V. Ranade (1989) J. Clin. Pharmacol. 29: 685). Exampletargeting moieties include folate or biotin (see, e.g., U.S. Pat. No.5,416,016 to Low et al.); mannosides (Umezawa et al., (1988) Biochem.Biophys. Res. Commun. 153: 1038); antibodies (P. G. Bloeman et al.(1995) FEBS Lett. 357: 140; M. Owais et al. (1995) Antimicrob. AgentsChemother. 39: 180); surfactant protein A receptor (Briscoe et al.(1995) Am. J. Physiol. 1233: 134), different species of which maycomprise the formulations of the disclosures, as well as components ofthe invented molecules; p120 (Schreier et al. (1994) J. Biol. Chem. 269:9090); see also K. Keinanen; M. L. Laukkanen (1994) FEBS Lett. 346: 123;J. J. Killion; I. J. Fidler (1994) Immunomethods 4: 273.

Particular Specific Embodiments

In a particular specific embodiment, the present disclosure is acomposition comprising one or more HSF1 RNAi agents.

In one embodiment, the disclosure comprises or consists of: AD-20278, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20279, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20280, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20281, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20282, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20283, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20303, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20313, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20315, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20348, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20362, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20364, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20365, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20366, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20373, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20376, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20377, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20378, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20386, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20389, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20391, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20392, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20397, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20398, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20399, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20401, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20402, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20403, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20404, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20406, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20407, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20408, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20409, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20410, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20411, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20413, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20422, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20428, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20434, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20435, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20437.4,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20437, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20438, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20439, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20487.7,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20487, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20488, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20489.2,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20489, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20490, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20491, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20493, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20495, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20502, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20507, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20513, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20527, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20535, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20544, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20545, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20546, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20547, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20548, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20549, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20552, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20555, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20556, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20557, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20558, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20560.4,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20560, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20561, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20562, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20563, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20564, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20565, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20566, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20570, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20572, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20574, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20575, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20577, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20578, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20579, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20580, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20597, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20598, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20625, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20626, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20627, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20633, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20634, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20640, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20644, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20646, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20648, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20650, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20652, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20653, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20660, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20661, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20671, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20693, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20694, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20700, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20702, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20707, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20709, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20710, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20714, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20716, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20728, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20730, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20741, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20764, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-20783, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-30071, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-36969, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-36970, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37718, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37719, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37720, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37721, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37722, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37723, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37724, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37725, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37726, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37727, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37728, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37729, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37730, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37731, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37732, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37733, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37734, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37735, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-30071.2,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-36969.2,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-36970.2,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37718.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37719.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37720.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37721.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37722.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37723.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37724.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37725.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37726.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37727.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37728.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37729.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37730.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37731.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37732.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37733.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37734.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37735.1,or modified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37736, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37737, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37738, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37739, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37740, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37741, ormodified or unmodified variants thereof.

In one embodiment, the disclosure comprises or consists of: AD-37742, ormodified or unmodified variants thereof.

Additional Particular Specific Embodiments

In various embodiments, the disclosure comprises a RNAi agent comprisinga first and a second strand, wherein the first strand comprises at least15 contiguous nucleotides differing by 0, 1, 2, or 3 nt from a firststrand, and the second strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from the second strand of anyone or more RNAi agent disclosed herein.

Various embodiments are further delineated below.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20278, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20279, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20280, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20281, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20282, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20283, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20303, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20313, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20315, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20348, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20362, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20364, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20365, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20366, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20373, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20376, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20377, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20378, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20386, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20389, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20391, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20392, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20397, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20398, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20399, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20401, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20402, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20403, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20404, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20406, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20407, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20408, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20409, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20410, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20411, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20413, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20422, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20428, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20434, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20435, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20437.4, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20437, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20438, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20439, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20487.7, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20487, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20488, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20489.2, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20489, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20490, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20491, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20493, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20495, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20502, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20507, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20513, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20527, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20535, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20544, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20545, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20546, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20547, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20548, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20549, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20552, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20555, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20556, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20557, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20558, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20560.4, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20560, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20561, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20562, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20563, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20564, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20565, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20566, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20570, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20572, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20574, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20575, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20577, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20578, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20579, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20580, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20597, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20598, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20625, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20626, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20627, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20633, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20634, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20640, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20644, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20646, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20648, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20650, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20652, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20653, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20660, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20661, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20671, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20693, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20694, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20700, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20702, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20707, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20709, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20710, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20714, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20716, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20728, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20730, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20741, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20764, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-20783, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-30071 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-36969 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-36970 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37718 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37719 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37720 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37721 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37722 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37723 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37724 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37725 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37726 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37727 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37728 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37729 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37730 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37731 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37732 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37733 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37734 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37735 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37736 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37737 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37738 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37739 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37740 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37741 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37742 or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-30071.2, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-36969.2, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-36970.2, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37718.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37719.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37720.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37721.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37722.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37723.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37724.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37725.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37726.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37727.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37728.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37729.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37730.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37731.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37732.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37733.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37734.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37735.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37736.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37737.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37738.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37739.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37740.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37741.1, or modified orunmodified variants thereof.

The disclosure comprises a RNAi agent comprising a first and a secondstrand, wherein the first strand comprises at least 15 contiguousnucleotides differing by 0, 1, 2, or 3 nt from a first strand, and thesecond strand comprises at least 15 contiguous nucleotides differing by0, 1, 2, or 3 nt from the second strand of: AD-37742.1, or modified orunmodified variants thereof.

Additional Particular Specific Embodiments

In various embodiments, the disclosure comprises a RNAi agent comprisinga sense and an antisense strand, wherein the antisense strand comprisesat least 15 contiguous nucleotides differing by 0, 1, 2, or 3 nt fromthe antisense strand of any RNAi agent disclosed herein.

Thus, in various embodiments:

The disclosure comprises a RNAi agent comprising a sense and anantisense strand, wherein the antisense strand comprises at least 15contiguous nucleotides differing by 0, 1, 2, or 3 nt from the antisensestrand of any one or more of the following duplexes, or modified orunmodified variants thereof: AD-20278, AD-20279, AD-20280, AD-20281,AD-20282, AD-20283, AD-20296, AD-20300, AD-20303, AD-20312, AD-20313,AD-20315, AD-20344, AD-20345, AD-20348, AD-20349, AD-20353, AD-20362,AD-20364, AD-20365, AD-20366, AD-20373, AD-20374, AD-20376, AD-20377,AD-20378, AD-20379, AD-20380, AD-20386, AD-20387, AD-20388, AD-20389,AD-20390, AD-20391, AD-20392, AD-20393, AD-20395, AD-20396, AD-20397,AD-20398, AD-20399, AD-20401 AD-20402, AD-20403, AD-20404, AD-20406,AD-20407, AD-20408, AD-20409, AD-20410, AD-20411, AD-20413, AD-20421,AD-20422, AD-20424, AD-20426, AD-20426, AD-20427, AD-20428, AD-20433,AD-20434, AD-20435, AD-20436, AD-20437, AD-20438, AD-20439, AD-20487,AD-20488, AD-20489, AD-20490, AD-20491, AD-20492, AD-20493, AD-20494,AD-20495, AD-20501, AD-20502, AD-20504, AD-20506, AD-20507, AD-20510,AD-20511, AD-20513, AD-20527, AD-20530 AD-20531, AD-20534, AD-20535,AD-20538, AD-20542, AD-20543, AD-20544, AD-20545, AD-20546, AD-20547,AD-20548, AD-20549, AD-20550, AD-20552, AD-20554, AD-20555, AD-20556,AD-20557, AD-20558, AD-20559, AD-20560, AD-20561, AD-20562, AD-20563,AD-20564, AD-20565, AD-20566, AD-20567, AD-20570, AD-20572, AD-20574,AD-20575, AD-20576, AD-20577, AD-20578, AD-20579, AD-20580, AD-20581,AD-20582, AD-20625, AD-20626, AD-20627, AD-20628, AD-20629, AD-20630,AD-20631, AD-20632, AD-20633, AD-20635, AD-20638, AD-20639, AD-20640,AD-20642, AD-20643, AD-20644, AD-20646, AD-20647, AD-20648, AD-20650,AD-20652, AD-20653, AD-20656, AD-20658, AD-20659, AD-20660, AD-20661,AD-20662, AD-20670, AD-20671, AD-20672, AD-20676, AD-20678, AD-20693,AD-20694, AD-20695, AD-20700, AD-20701, AD-20702, AD-20703, AD-20705,AD-20706, AD-20707, AD-20708, AD-20709, AD-20710, AD-20711, AD-20713,AD-20714, AD-20715, AD-20716, AD-20718, AD-20720, AD-20728, AD-20730,AD-20731, AD-20741, AD-20742, AD-20743, AD-20744, AD-20748, AD-20751,AD-20752, AD-20754, AD-20764, AD-20765, AD-20766, AD-20783, AD-20784,AD-20785, AD-20786, AD-20790, AD-20801, or modified or unmodifiedvariants thereof.

Additional Particular Embodiments

In various embodiments, the disclosure comprises a RNAi agent comprisinga sense and an antisense strand, wherein the antisense strand comprisesor consists of the antisense strand of any RNAi agent disclosed herein.

Thus, the following are provided as examples of the various embodiments.

The disclosure comprises a RNAi agent comprising a sense and anantisense strand, wherein the antisense strand comprises or consists ofthe antisense strand of: AD-20278, AD-20279, AD-20280, AD-20281,AD-20282, AD-20283, AD-20296, AD-20300, AD-20303, AD-20312, AD-20313,AD-20315, AD-20344, AD-20345, AD-20348, AD-20349, AD-20353, AD-20362,AD-20364, AD-20365, AD-20366, AD-20373, AD-20374, AD-20376, AD-20377,AD-20378, AD-20379, AD-20380, AD-20386, AD-20387, AD-20388, AD-20389,AD-20390, AD-20391, AD-20392, AD-20393, AD-20395, AD-20396, AD-20397,AD-20398, AD-20399, AD-20401 AD-20402, AD-20403, AD-20404, AD-20406,AD-20407, AD-20408, AD-20409, AD-20410, AD-20411, AD-20413, AD-20421,AD-20422, AD-20424, AD-20426, AD-20426, AD-20427, AD-20428, AD-20433,AD-20434, AD-20435, AD-20436, AD-20437, AD-20438, AD-20439, AD-20487,AD-20488, AD-20489, AD-20490, AD-20491, AD-20492, AD-20493, AD-20494,AD-20495, AD-20501, AD-20502, AD-20504, AD-20506, AD-20507, AD-20510,AD-20511, AD-20513, AD-20527, AD-20530 AD-20531, AD-20534, AD-20535,AD-20538, AD-20542, AD-20543, AD-20544, AD-20545, AD-20546, AD-20547,AD-20548, AD-20549, AD-20550, AD-20552, AD-20554, AD-20555, AD-20556,AD-20557, AD-20558, AD-20559, AD-20560, AD-20561, AD-20562, AD-20563,AD-20564, AD-20565, AD-20566, AD-20567, AD-20570, AD-20572, AD-20574,AD-20575, AD-20576, AD-20577, AD-20578, AD-20579, AD-20580, AD-20581,AD-20582, AD-20625, AD-20626, AD-20627, AD-20628, AD-20629, AD-20630,AD-20631, AD-20632, AD-20633, AD-20635, AD-20638, AD-20639, AD-20640,AD-20642, AD-20643, AD-20644, AD-20646, AD-20647, AD-20648, AD-20650,AD-20652, AD-20653, AD-20656, AD-20658, AD-20659, AD-20660, AD-20661,AD-20662, AD-20670, AD-20671, AD-20672, AD-20676, AD-20678, AD-20693,AD-20694, AD-20695, AD-20700, AD-20701, AD-20702, AD-20703, AD-20705,AD-20706, AD-20707, AD-20708, AD-20709, AD-20710, AD-20711, AD-20713,AD-20714, AD-20715, AD-20716, AD-20718, AD-20720, AD-20728, AD-20730,AD-20731, AD-20741, AD-20742, AD-20743, AD-20744, AD-20748, AD-20751,AD-20752, AD-20754, AD-20764, AD-20765, AD-20766, AD-20783, AD-20784,AD-20785, AD-20786, AD-20790, AD-20801, or modified or unmodifiedvariants thereof.

In various embodiments, the disclosure comprises a RNAi agent comprisinga sense and an antisense strand, wherein the antisense strand comprisesat least 15 contiguous nucleotides differing by 0, 1, 2, or 3 nt fromthe antisense strand of any RNAi agent disclosed herein, or modified orunmodified variants thereof, wherein the antisense strand optionallyfurther comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more nt (or anyrange thereof, e.g., 0-1, 1-2, 1-3, 1-4 nt, etc.).

Thus, in various embodiments, the disclosure comprises a RNAi agentcomprising a sense and an antisense strand, wherein the antisense strandcomprises at least 15 contiguous nucleotides differing by 0, 1, 2, or 3nt from the antisense strand of: AD-20278, AD-20279, AD-20280, AD-20281,AD-20282, AD-20283, AD-20303, AD-20313, AD-20315, AD-20348, AD-20362,AD-20364, AD-20365, AD-20366, AD-20373, AD-20376, AD-20377, AD-20378,AD-20386, AD-20389, AD-20391, AD-20392, AD-20397, AD-20398, AD-20399,AD-20401, AD-20402, AD-20403, AD-20404, AD-20406, AD-20407, AD-20408,AD-20409, AD-20410, AD-20411, AD-20413, AD-20422, AD-20428, AD-20434,AD-20435, AD-20437, AD-20437, AD-20438, AD-20439, AD-20487, AD-20487,AD-20488, AD-20489, AD-20489, AD-20490, AD-20491, AD-20493, AD-20495,AD-20502, AD-20507, AD-20513, AD-20527, AD-20535, AD-20544, AD-20545,AD-20546, AD-20547, AD-20548, AD-20549, AD-20552, AD-20555, AD-20556,AD-20557, AD-20558, AD-20560, AD-20560, AD-20561, AD-20562, AD-20563,AD-20564, AD-20565, AD-20566, AD-20570, AD-20572, AD-20574, AD-20575,AD-20577, AD-20578, AD-20579, AD-20580, AD-20597, AD-20598, AD-20625,AD-20626, AD-20627, AD-20633, AD-20634, AD-20640, AD-20644, AD-20646,AD-20648, AD-20650, AD-20652, AD-20653, AD-20660, AD-20661, AD-20671,AD-20693, AD-20694, AD-20700, AD-20702, AD-20707, AD-20709, AD-20710,AD-20714, AD-20716, AD-20728, AD-20730, AD-20741, AD-20764, AD-20783,AD-30071, AD-36969, AD-36970, AD-37718, AD-37719, AD-37720, AD-37721,AD-37722, AD-37723, AD-37724, AD-37725, AD-37726, AD-37727, AD-37728,AD-37729, AD-37730, AD-37731, AD-37732, AD-37733, AD-37734, AD-37735,AD-37736, AD-37737, AD-37738, AD-37739, AD-37740, AD-37741, AD-37742, ormodified or unmodified variants thereof, wherein the antisense strandoptionally further comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or morent (or any range thereof, e.g., 0-1, 1-2, 1-3, 1-4 nt, etc.).

In one embodiment, the disclosure comprises any one or more RNAi agentlisted herein.

Additional Particular Specific Embodiments

Certain RNAi agents to HSF1 are disclosed in the scientific literature,e.g., in Rossi et al. 2006 Cancer Res. 66:7678-85; Dokladny et al. 2008Am. J. Pathology 72:659-70; Jacobs et al. 2007 J. Biol. Chem. 282:33412-20; Page et al. 2006 Mol. Biosystems 2:627-39; Zhao et al. 2007Diabetes 56: 1436-1444; and Du et al. 2009 J. Cell. Phys. 218:631-637.The compositions of this invention do not cover these RNAi agents to theextent that they are identical in both sequence and modifications.

Other particular specific embodiments include compositions comprising 1,2, 3, 4, or more of these RNAi agents. Another embodiment is acomposition comprising any single RNAi agent, along with any other RNAiagents which overlap it. Another embodiment comprises two, three, fouror more HSF1 RNAi agents which do not overlap and thus target differentparts of the RNA molecule. When two or more RNAi agents are used, theycan be administered simultaneously or sequentially.

Another particular specific embodiment comprises an RNAi agent, whereinthe

RNAi agent comprises a sense strand comprising at least 15 contiguousnucleotides (identical in sequence) to the sense strand of any of thelisted RNAi agents, and an antisense strand comprising at least 15contiguous nucleotides (identical in sequence) to the antisense strandof the same RNAi agent. In another embodiment, the composition comprisesone, two, three, four, or more such RNAi agents.

In one embodiment, the composition comprises an RNAi agent whichcomprises an antisense strand comprising at least 15 contiguousnucleotides differing by 0, 1, 2 or 3 mismatches from the antisensestrand of a RNAi agent described herein.

In one embodiment, the composition comprises an RNAi agent whichcomprises an antisense strand comprising at least 15 contiguousnucleotides differing by 0, 1, 2 or 3 mismatches from the antisensestrand of a RNAi agent described herein.

In another embodiment, the composition comprises an RNAi agent whichcomprises a sense strand comprising at least 15 contiguous nucleotidesdiffering by 0, 1, 2, or 3 mismatches from the sense strand of one ofthe listed RNAi agents, and an antisense strand comprising at least 15contiguous nucleotides differing by 0, 1, 2 or 3 mismatches from theantisense strand of the same RNAi agent.

A “mismatch” is defined herein as a difference between the base sequenceor length when two sequences are maximally aligned and compared. As anon-limiting example, a mismatch is counted if a difference existsbetween the base at a particular location in one sequence and the baseat the corresponding position in another sequence (e.g., between thesequence of a given RNAi agent and an RNAi agent listed herein). Thus, amismatch is counted, for example, if a position in one sequence has aparticular base (e.g., A), and the corresponding position on the othersequence has a different base (e.g., G, C or U). A mismatch is alsocounted, e.g., if a position in one sequence has a base (e.g., A), andthe corresponding position on the other sequence has no base (e.g., thatposition is an abasic nucleotide which comprises a phosphate-sugarbackbone but no base). A single-stranded nick in either sequence (or inthe sense or antisense strand) is not counted as mismatch. Thus, as anon-limiting example, no mismatch would be counted if one sequencecomprises the sequence A-G, but the other sequence comprises thesequence A-G with a single-stranded nick between the A and the G. A basemodification is also not considered a mismatch. Hone sequence comprisesa C, and the other sequence comprises a modified C (e.g., with a2′-modification) at the same position, no mismatch would be counted.Thus, modifications of a nucleotide other than replacement or alterationof the base would not constitute a mismatch. For example, no mismatchwould occur between a nucleotide which is A, and a nucleotide which is Awith a 5′ modification (e.g., those illustrated in FIG. 1) and/or a2′-modification. The key feature of a mismatch (base replacement) isthat it would not be able to base-pair with the corresponding base onthe opposite strand. In addition, terminal overhangs such as “UU” or“dTdT” are not counted when counting the number of mismatches; theterminal “UU” and “dTdT” overhangs are also not included whencalculating “15 contiguous nucleotides.”

In these embodiments, a mismatch is defined as a position wherein thebase of one sequence does not match the base of the other sequence.

In another embodiment, the composition comprises 1, 2, 3, 4, or moresuch RNAi agents.

In another embodiment, the composition comprises an RNAi agent whichcomprises a sense strand comprising at least 15 contiguous nucleotidesdiffering by 0, 1, 2 or 3 mismatches from the sense strand of one of thelisted RNAi agents, and an antisense strand comprising at least 15contiguous nucleotides differing by 0, 1, 2 or 3 mismatches from theantisense strand of the same RNAi agent

Overlapping Groups of HSF1 siRNAs

In various embodiments, the disclosure relates to groups of RNAi agentswith overlapping sequences. Thus, the disclosure encompasses groups ofRNAi agents wherein each RNAi agent in the group overlaps with eachother RNAi agent in the same group by at least 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19 or more nucleotides. Particularly, in oneembodiment, the overlap is at least 12 nt. Groups of sequences thatoverlap are shown in Table 3A.

Table 3A shows, for example, that AD-20594 and AD-20596 share the commontechnical feature of the sequence of ACGUCCCGGCCU in the sense strand,and the sequence of AGGCCGGGACGU in the antisense strand. Note of coursethat only a 12-nt portion of the overlap is shown; many groups of RNAiagents will overlap by more than 12 nt. The position within the gene isalso indicated.

The disclosure thus encompasses various embodiments comprising groups ofoverlapping RNAi agents, for example (1) RNAi agents comprising thesequences of AD-20594 and AD-20596; (2) RNAi agents consisting of thesequences of AD-20594 and AD-20596; (3) RNAi agents comprising thesequences of AD-20594 and AD-20596; (4) RNAi agents comprising a sensestrand and/or a antisense strand comprising a sequence of AD-20594 andAD-20596; (5) RNAi agents comprising a sense strand and/or a antisensestrand comprising 15 contiguous nt with 0 to 3 mismatches from asequence of AD-20594 and AD-20596; (6) RNAi agents comprising a sensestrand comprising 15 contiguous nt with 0 to 3 mismatches from asequence of AD-20594 and AD-20596; (7) RNAi agents comprising anantisense strand comprising 15 contiguous nt with 0 to 3 mismatches froma sequence of AD-20594 and AD-20596; etc. The disclosure alsoencompasses similar embodiments reflecting all the overlapping groups ofRNAi agents as described in Table 3A.

Variants of RNAi agents (e.g., comprising different modifications, caps,etc.) are disclosed herein, e.g., in Tables 2, 3, 9A and 9B. In thesetexts and tables, for example, AD-20437 shares the same sequence asAD-20437.4, though the RNAi agents differ in their modifications, caps(e.g., 5′ and/or 3′ caps), etc. However, any overlapping groupcomprising a RNAi agent of a given sequence also comprises any otherRNAi agent which has the same sequence, but different variations inmodifications, caps, etc. Thus, any group of overlapping RNAi agentsthat includes AD-20437 also includes AD-20437.4 and other variants ofthe same sequence (e.g., with different modifications, caps, etc.). Moreembodiments are provided herein, and are included in the scope of eachRNAi agents of the disclosure.

EXAMPLES Example 1 Bioinformatics

Transcripts

Oligonucleotide design was carried out to identify siRNAs targeting thegene “heat shock transcription factor 1 (HSF1)” from human (NCBI symbolHSF1), the orthologous sequences from rhesus monkey (Macaca mulatta),cynomolgus monkey (Macaca fascicularis), and orangutan (Pongo pygmaeus).The design process used the HSF1 transcripts NM_(—)005526.2 from human(NCBI Geneld 3297), ENSMMUT00000020648 from rhesus (from Ensembl),internally cloned and sequenced cyno HSF1 sequences (e.g., SEQ ID NO:2051), and ENSPPYT00000022122 from orangutan (from Ensembl). The designstrategy was as follows: Begin with all perfect 19-mer human/cyno HSF1matches. Next, expand this set with all perfect 19-merhuman/orangutan/rhesus matches. Any 19-mers with mismatches to thepartial cyno sequence available at the time of design were thenexcluded. Last, seven sequences were selected based on homology with theNovartis shRNA library. The resulting set of 512 19-mer sequences, allperfect matches to the human HSF1 gene, were then further sampled forsynthesis and screening. These sequences are listed in Table 1. Thesesequences have not been modified.

The phrase “Position” denotes the starting position of theoligonucleotide (e.g., the 19-mer) on the transcript. This is measuredin nucleotide coordinates, relative to beginning of the transcript.

TABLE 1 HSF1 19-mers Duplex SEQ SEQ ID Name Position ID NO Sense5′-3′ unmodified NO Antisense 5′-3′ unmodified R0001 201 1GGGCCCAGCAACGUCCCGG 513 CCGGGACGUUGCUGGGCCC R0002 202 2GGCCCAGCAACGUCCCGGC 514 GCCGGGACGUUGCUGGGCC R0003 203 3GCCCAGCAACGUCCCGGCC 515 GGCCGGGACGUUGCUGGGC R0004 204 4CCCAGCAACGUCCCGGCCU 516 AGGCCGGGACGUUGCUGGG R0005 205 5CCAGCAACGUCCCGGCCUU 517 AAGGCCGGGACGUUGCUGG R0006 206 6CAGCAACGUCCCGGCCUUC 518 GAAGGCCGGGACGUUGCUG R0007 207 7AGCAACGUCCCGGCCUUCC 519 GGAAGGCCGGGACGUUGCU R0008 208 8GCAACGUCCCGGCCUUCCU 520 AGGAAGGCCGGGACGUUGC AD-20594 209 9CAACGUCCCGGCCUUCCUG 521 CAGGAAGGCCGGGACGUUG AD-20595 210 10AACGUCCCGGCCUUCCUGA 522 UCAGGAAGGCCGGGACGUU AD-20596 211 11ACGUCCCGGCCUUCCUGAC 523 GUCAGGAAGGCCGGGACGU AD-20285 212 12CGUCCCGGCCUUCCUGACC 524 GGUCAGGAAGGCCGGGACG AD-20286 213 13GUCCCGGCCUUCCUGACCA 525 UGGUCAGGAAGGCCGGGAC AD-20287 216 14CCGGCCUUCCUGACCAAGC 526 GCUUGGUCAGGAAGGCCGG AD-20288 217 15CGGCCUUCCUGACCAAGCU 527 AGCUUGGUCAGGAAGGCCG AD-20289 218 16GGCCUUCCUGACCAAGCUG 528 CAGCUUGGUCAGGAAGGCC AD-20290 219 17GCCUUCCUGACCAAGCUGU 529 ACAGCUUGGUCAGGAAGGC AD-20291 220 18CCUUCCUGACCAAGCUGUG 530 CACAGCUUGGUCAGGAAGG AD-20292 221 19CUUCCUGACCAAGCUGUGG 531 CCACAGCUUGGUCAGGAAG AD-20293 222 20UUCCUGACCAAGCUGUGGA 532 UCCACAGCUUGGUCAGGAA AD-20294 223 21UCCUGACCAAGCUGUGGAC 533 GUCCACAGCUUGGUCAGGA AD-20295 224 22CCUGACCAAGCUGUGGACC 534 GGUCCACAGCUUGGUCAGG AD-20296 225 23CUGACCAAGCUGUGGACCC 535 GGGUCCACAGCUUGGUCAG AD-20297 226 24UGACCAAGCUGUGGACCCU 536 AGGGUCCACAGCUUGGUCA AD-20298 227 25GACCAAGCUGUGGACCCUC 537 GAGGGUCCACAGCUUGGUC AD-20299 228 26ACCAAGCUGUGGACCCUCG 538 CGAGGGUCCACAGCUUGGU AD-20300 229 27CCAAGCUGUGGACCCUCGU 539 ACGAGGGUCCACAGCUUGG AD-20301 230 28CAAGCUGUGGACCCUCGUG 540 CACGAGGGUCCACAGCUUG AD-20302 231 29AAGCUGUGGACCCUCGUGA 541 UCACGAGGGUCCACAGCUU AD-20303 232 30AGCUGUGGACCCUCGUGAG 542 CUCACGAGGGUCCACAGCU AD-20304 233 31GCUGUGGACCCUCGUGAGC 543 GCUCACGAGGGUCCACAGC AD-20305 234 32CUGUGGACCCUCGUGAGCG 544 CGCUCACGAGGGUCCACAG AD-20306 235 33UGUGGACCCUCGUGAGCGA 545 UCGCUCACGAGGGUCCACA AD-20307 236 34GUGGACCCUCGUGAGCGAC 546 GUCGCUCACGAGGGUCCAC AD-20308 237 35UGGACCCUCGUGAGCGACC 547 GGUCGCUCACGAGGGUCCA AD-20309 238 36GGACCCUCGUGAGCGACCC 548 GGGUCGCUCACGAGGGUCC AD-20310 239 37GACCCUCGUGAGCGACCCG 549 CGGGUCGCUCACGAGGGUC AD-20311 240 38ACCCUCGUGAGCGACCCGG 550 CCGGGUCGCUCACGAGGGU AD-20312 241 39CCCUCGUGAGCGACCCGGA 551 UCCGGGUCGCUCACGAGGG AD-20313 242 40CCUCGUGAGCGACCCGGAC 552 GUCCGGGUCGCUCACGAGG AD-20314 243 41CUCGUGAGCGACCCGGACA 553 UGUCCGGGUCGCUCACGAG AD-20315 244 42UCGUGAGCGACCCGGACAC 554 GUGUCCGGGUCGCUCACGA AD-20316 245 43CGUGAGCGACCCGGACACC 555 GGUGUCCGGGUCGCUCACG AD-20317 246 44GUGAGCGACCCGGACACCG 556 CGGUGUCCGGGUCGCUCAC AD-20318 247 45UGAGCGACCCGGACACCGA 557 UCGGUGUCCGGGUCGCUCA R0009 248 46GAGCGACCCGGACACCGAC 558 GUCGGUGUCCGGGUCGCUC R0010 249 47AGCGACCCGGACACCGACG 559 CGUCGGUGUCCGGGUCGCU R0011 250 48GCGACCCGGACACCGACGC 560 GCGUCGGUGUCCGGGUCGC AD-20319 270 49CUCAUCUGCUGGAGCCCGA 561 UCGGGCUCCAGCAGAUGAG AD-20320 271 50UCAUCUGCUGGAGCCCGAG 562 CUCGGGCUCCAGCAGAUGA AD-20344 306 51GUGUUCGACCAGGGCCAGU 563 ACUGGCCCUGGUCGAACAC AD-20345 307 52UGUUCGACCAGGGCCAGUU 564 AACUGGCCCUGGUCGAACA R0103 308 53GUUCGACCAGGGCCAGUUU 565 AAACUGGCCCUGGUCGAAC AD-20346 309 54UUCGACCAGGGCCAGUUUG 566 CAAACUGGCCCUGGUCGAA AD-20347 310 55UCGACCAGGGCCAGUUUGC 567 GCAAACUGGCCCUGGUCGA AD-20348 311 56CGACCAGGGCCAGUUUGCC 568 GGCAAACUGGCCCUGGUCG AD-20349 312 57GACCAGGGCCAGUUUGCCA 569 UGGCAAACUGGCCCUGGUC AD-20350 313 58ACCAGGGCCAGUUUGCCAA 570 UUGGCAAACUGGCCCUGGU AD-20351 314 59CCAGGGCCAGUUUGCCAAG 571 CUUGGCAAACUGGCCCUGG AD-20352 315 60CAGGGCCAGUUUGCCAAGG 572 CCUUGGCAAACUGGCCCUG AD-20353 316 61AGGGCCAGUUUGCCAAGGA 573 UCCUUGGCAAACUGGCCCU AD-20354 317 62GGGCCAGUUUGCCAAGGAG 574 CUCCUUGGCAAACUGGCCC AD-20355 318 63GGCCAGUUUGCCAAGGAGG 575 CCUCCUUGGCAAACUGGCC AD-20356 319 64GCCAGUUUGCCAAGGAGGU 576 ACCUCCUUGGCAAACUGGC AD-20357 320 65CCAGUUUGCCAAGGAGGUG 577 CACCUCCUUGGCAAACUGG AD-20358 321 66CAGUUUGCCAAGGAGGUGC 578 GCACCUCCUUGGCAAACUG AD-20359 322 67AGUUUGCCAAGGAGGUGCU 579 AGCACCUCCUUGGCAAACU AD-20360 323 68GUUUGCCAAGGAGGUGCUG 580 CAGCACCUCCUUGGCAAAC AD-20361 324 69UUUGCCAAGGAGGUGCUGC 581 GCAGCACCUCCUUGGCAAA AD-20362 325 70UUGCCAAGGAGGUGCUGCC 582 GGCAGCACCUCCUUGGCAA AD-20363 326 71UGCCAAGGAGGUGCUGCCC 583 GGGCAGCACCUCCUUGGCA AD-20364 327 72GCCAAGGAGGUGCUGCCCA 584 UGGGCAGCACCUCCUUGGC AD-20365 328 73CCAAGGAGGUGCUGCCCAA 585 UUGGGCAGCACCUCCUUGG AD-20366 329 74CAAGGAGGUGCUGCCCAAG 586 CUUGGGCAGCACCUCCUUG AD-20367 330 75AAGGAGGUGCUGCCCAAGU 587 ACUUGGGCAGCACCUCCUU AD-20368 331 76AGGAGGUGCUGCCCAAGUA 588 UACUUGGGCAGCACCUCCU AD-20369 351 77UUCAAGCACAACAACAUGG 589 CCAUGUUGUUGUGCUUGAA AD-20370 352 78UCAAGCACAACAACAUGGC 590 GCCAUGUUGUUGUGCUUGA AD-20371 353 79CAAGCACAACAACAUGGCC 591 GGCCAUGUUGUUGUGCUUG AD-20372 354 80AAGCACAACAACAUGGCCA 592 UGGCCAUGUUGUUGUGCUU AD-20373 355 81AGCACAACAACAUGGCCAG 593 CUGGCCAUGUUGUUGUGCU AD-20374 356 82GCACAACAACAUGGCCAGC 594 GCUGGCCAUGUUGUUGUGC AD-20375 357 83CACAACAACAUGGCCAGCU 595 AGCUGGCCAUGUUGUUGUG AD-20376 358 84ACAACAACAUGGCCAGCUU 596 AAGCUGGCCAUGUUGUUGU AD-20378 360 85AACAACAUGGCCAGCUUCG 597 CGAAGCUGGCCAUGUUGUU AD-20379 361 86ACAACAUGGCCAGCUUCGU 598 ACGAAGCUGGCCAUGUUGU AD-20380 362 87CAACAUGGCCAGCUUCGUG 599 CACGAAGCUGGCCAUGUUG AD-20381 363 88AACAUGGCCAGCUUCGUGC 600 GCACGAAGCUGGCCAUGUU AD-20382 364 89ACAUGGCCAGCUUCGUGCG 601 CGCACGAAGCUGGCCAUGU AD-20383 365 90CAUGGCCAGCUUCGUGCGG 602 CCGCACGAAGCUGGCCAUG AD-20384 366 91AUGGCCAGCUUCGUGCGGC 603 GCCGCACGAAGCUGGCCAU AD-20385 367 92UGGCCAGCUUCGUGCGGCA 604 UGCCGCACGAAGCUGGCCA AD-20386 436 93UGGUCAAGCCAGAGAGAGA 605 UCUCUCUCUGGCUUGACCA R0012 437 94GGUCAAGCCAGAGAGAGAC 606 GUCUCUCUCUGGCUUGACC R0013 438 95GUCAAGCCAGAGAGAGACG 607 CGUCUCUCUCUGGCUUGAC R0014 439 96UCAAGCCAGAGAGAGACGA 608 UCGUCUCUCUCUGGCUUGA R0015 440 97CAAGCCAGAGAGAGACGAC 609 GUCGUCUCUCUCUGGCUUG R0016 441 98AAGCCAGAGAGAGACGACA 610 UGUCGUCUCUCUCUGGCUU R0017 442 99AGCCAGAGAGAGACGACAC 611 GUGUCGUCUCUCUCUGGCU R0018 443 100GCCAGAGAGAGACGACACG 612 CGUGUCGUCUCUCUCUGGC R0019 444 101CCAGAGAGAGACGACACGG 613 CCGUGUCGUCUCUCUCUGG R0020 445 102CAGAGAGAGACGACACGGA 614 UCCGUGUCGUCUCUCUCUG R0021 446 103AGAGAGAGACGACACGGAG 615 CUCCGUGUCGUCUCUCUCU R0022 447 104GAGAGAGACGACACGGAGU 616 ACUCCGUGUCGUCUCUCUC R0023 448 105AGAGAGACGACACGGAGUU 617 AACUCCGUGUCGUCUCUCU R0024 449 106GAGAGACGACACGGAGUUC 618 GAACUCCGUGUCGUCUCUC R0025 450 107AGAGACGACACGGAGUUCC 619 GGAACUCCGUGUCGUCUCU R0026 451 108GAGACGACACGGAGUUCCA 620 UGGAACUCCGUGUCGUCUC R0027 452 109AGACGACACGGAGUUCCAG 621 CUGGAACUCCGUGUCGUCU R0028 453 110GACGACACGGAGUUCCAGC 622 GCUGGAACUCCGUGUCGUC R0029 454 111ACGACACGGAGUUCCAGCA 623 UGCUGGAACUCCGUGUCGU R0030 455 112CGACACGGAGUUCCAGCAC 624 GUGCUGGAACUCCGUGUCG R0031 456 113GACACGGAGUUCCAGCACC 625 GGUGCUGGAACUCCGUGUC R0032 457 114ACACGGAGUUCCAGCACCC 626 GGGUGCUGGAACUCCGUGU AD-20387 489 115GGCCAGGAGCAGCUCCUUG 627 CAAGGAGCUGCUCCUGGCC AD-20388 490 116GCCAGGAGCAGCUCCUUGA 628 UCAAGGAGCUGCUCCUGGC AD-20389 491 117CCAGGAGCAGCUCCUUGAG 629 CUCAAGGAGCUGCUCCUGG AD-20390 492 118CAGGAGCAGCUCCUUGAGA 630 UCUCAAGGAGCUGCUCCUG AD-20391 493 119AGGAGCAGCUCCUUGAGAA 631 UUCUCAAGGAGCUGCUCCU AD-20392 494 120GGAGCAGCUCCUUGAGAAC 632 GUUCUCAAGGAGCUGCUCC AD-20393 495 121GAGCAGCUCCUUGAGAACA 633 UGUUCUCAAGGAGCUGCUC AD-20394 496 122AGCAGCUCCUUGAGAACAU 634 AUGUUCUCAAGGAGCUGCU AD-20395 497 123GCAGCUCCUUGAGAACAUC 635 GAUGUUCUCAAGGAGCUGC AD-20396 498 124CAGCUCCUUGAGAACAUCA 636 UGAUGUUCUCAAGGAGCUG AD-20397 499 125AGCUCCUUGAGAACAUCAA 637 UUGAUGUUCUCAAGGAGCU AD-20398 500 126GCUCCUUGAGAACAUCAAG 638 CUUGAUGUUCUCAAGGAGC AD-20399 501 127CUCCUUGAGAACAUCAAGA 639 UCUUGAUGUUCUCAAGGAG AD-20400 502 128UCCUUGAGAACAUCAAGAG 640 CUCUUGAUGUUCUCAAGGA AD-20401 503 129CCUUGAGAACAUCAAGAGG 641 CCUCUUGAUGUUCUCAAGG AD-20402 504 130CUUGAGAACAUCAAGAGGA 642 UCCUCUUGAUGUUCUCAAG AD-20403 505 131UUGAGAACAUCAAGAGGAA 643 UUCCUCUUGAUGUUCUCAA AD-20404 506 132UGAGAACAUCAAGAGGAAA 644 UUUCCUCUUGAUGUUCUCA R0033 507 133GAGAACAUCAAGAGGAAAG 645 CUUUCCUCUUGAUGUUCUC R0034 508 134AGAACAUCAAGAGGAAAGU 646 ACUUUCCUCUUGAUGUUCU AD-20405 509 135GAACAUCAAGAGGAAAGUG 647 CACUUUCCUCUUGAUGUUC AD-20406 510 136AACAUCAAGAGGAAAGUGA 648 UCACUUUCCUCUUGAUGUU AD-20407 511 137ACAUCAAGAGGAAAGUGAC 649 GUCACUUUCCUCUUGAUGU AD-20408 512 138CAUCAAGAGGAAAGUGACC 650 GGUCACUUUCCUCUUGAUG AD-20409 513 139AUCAAGAGGAAAGUGACCA 651 UGGUCACUUUCCUCUUGAU AD-20410 514 140UCAAGAGGAAAGUGACCAG 652 CUGGUCACUUUCCUCUUGA AD-20411 515 141CAAGAGGAAAGUGACCAGU 653 ACUGGUCACUUUCCUCUUG AD-20412 516 142AAGAGGAAAGUGACCAGUG 654 CACUGGUCACUUUCCUCUU AD-20413 517 2042AGAGGAAAGUGACCAGUGU 2043 ACACUGGUCACUUUCCUCU AD-20414 518 143GAGGAAAGUGACCAGUGUG 655 CACACUGGUCACUUUCCUC AD-20415 519 144AGGAAAGUGACCAGUGUGU 656 ACACACUGGUCACUUUCCU AD-20416 520 145GGAAAGUGACCAGUGUGUC 657 GACACACUGGUCACUUUCC AD-20417 521 146GAAAGUGACCAGUGUGUCC 658 GGACACACUGGUCACUUUC AD-20418 522 147AAAGUGACCAGUGUGUCCA 659 UGGACACACUGGUCACUUU AD-20419 523 148AAGUGACCAGUGUGUCCAC 660 GUGGACACACUGGUCACUU AD-20420 524 149AGUGACCAGUGUGUCCACC 661 GGUGGACACACUGGUCACU AD-20421 525 150GUGACCAGUGUGUCCACCC 662 GGGUGGACACACUGGUCAC AD-20422 526 151UGACCAGUGUGUCCACCCU 663 AGGGUGGACACACUGGUCA AD-20423 527 152GACCAGUGUGUCCACCCUG 664 CAGGGUGGACACACUGGUC AD-20424 528 153ACCAGUGUGUCCACCCUGA 665 UCAGGGUGGACACACUGGU AD-20425 529 154CCAGUGUGUCCACCCUGAA 666 UUCAGGGUGGACACACUGG AD-20426 530 155CAGUGUGUCCACCCUGAAG 667 CUUCAGGGUGGACACACUG AD-20427 531 156AGUGUGUCCACCCUGAAGA 668 UCUUCAGGGUGGACACACU AD-20428 532 157GUGUGUCCACCCUGAAGAG 669 CUCUUCAGGGUGGACACAC AD-20429 533 158UGUGUCCACCCUGAAGAGU 670 ACUCUUCAGGGUGGACACA AD-20430 534 159GUGUCCACCCUGAAGAGUG 671 CACUCUUCAGGGUGGACAC AD-20431 535 160UGUCCACCCUGAAGAGUGA 672 UCACUCUUCAGGGUGGACA AD-20432 536 161GUCCACCCUGAAGAGUGAA 673 UUCACUCUUCAGGGUGGAC AD-20433 537 162UCCACCCUGAAGAGUGAAG 674 CUUCACUCUUCAGGGUGGA AD-20434 538 163CCACCCUGAAGAGUGAAGA 675 UCUUCACUCUUCAGGGUGG AD-20435 539 164CACCCUGAAGAGUGAAGAC 676 GUCUUCACUCUUCAGGGUG AD-20436 540 165ACCCUGAAGAGUGAAGACA 677 UGUCUUCACUCUUCAGGGU AD-20437 541 166CCCUGAAGAGUGAAGACAU 678 AUGUCUUCACUCUUCAGGG AD-20438 542 167CCUGAAGAGUGAAGACAUA 679 UAUGUCUUCACUCUUCAGG AD-20439 543 168CUGAAGAGUGAAGACAUAA 680 UUAUGUCUUCACUCUUCAG AD-20487 544 169UGAAGAGUGAAGACAUAAA 681 UUUAUGUCUUCACUCUUCA AD-20488 545 170GAAGAGUGAAGACAUAAAG 682 CUUUAUGUCUUCACUCUUC AD-20489 546 171AAGAGUGAAGACAUAAAGA 683 UCUUUAUGUCUUCACUCUU AD-20490 547 172AGAGUGAAGACAUAAAGAU 684 AUCUUUAUGUCUUCACUCU AD-20491 548 173GAGUGAAGACAUAAAGAUC 685 GAUCUUUAUGUCUUCACUC AD-20492 549 174AGUGAAGACAUAAAGAUCC 686 GGAUCUUUAUGUCUUCACU AD-20493 550 175GUGAAGACAUAAAGAUCCG 687 CGGAUCUUUAUGUCUUCAC AD-20494 579 176GUCACCAAGCUGCUGACGG 688 CCGUCAGCAGCUUGGUGAC AD-20495 580 177UCACCAAGCUGCUGACGGA 689 UCCGUCAGCAGCUUGGUGA AD-20496 581 178CACCAAGCUGCUGACGGAC 690 GUCCGUCAGCAGCUUGGUG AD-20497 582 179ACCAAGCUGCUGACGGACG 691 CGUCCGUCAGCAGCUUGGU AD-20498 583 180CCAAGCUGCUGACGGACGU 692 ACGUCCGUCAGCAGCUUGG AD-20499 584 181CAAGCUGCUGACGGACGUG 693 CACGUCCGUCAGCAGCUUG AD-20500 585 182AAGCUGCUGACGGACGUGC 694 GCACGUCCGUCAGCAGCUU AD-20501 586 183AGCUGCUGACGGACGUGCA 695 UGCACGUCCGUCAGCAGCU AD-20502 587 184GCUGCUGACGGACGUGCAG 696 CUGCACGUCCGUCAGCAGC AD-20503 588 185CUGCUGACGGACGUGCAGC 697 GCUGCACGUCCGUCAGCAG AD-20504 589 186UGCUGACGGACGUGCAGCU 698 AGCUGCACGUCCGUCAGCA AD-20505 590 187GCUGACGGACGUGCAGCUG 699 CAGCUGCACGUCCGUCAGC AD-20506 591 188CUGACGGACGUGCAGCUGA 700 UCAGCUGCACGUCCGUCAG AD-20507 592 189UGACGGACGUGCAGCUGAU 701 AUCAGCUGCACGUCCGUCA AD-20508 593 190GACGGACGUGCAGCUGAUG 702 CAUCAGCUGCACGUCCGUC AD-20509 594 191ACGGACGUGCAGCUGAUGA 703 UCAUCAGCUGCACGUCCGU AD-20510 595 192CGGACGUGCAGCUGAUGAA 704 UUCAUCAGCUGCACGUCCG AD-20511 596 193GGACGUGCAGCUGAUGAAG 705 CUUCAUCAGCUGCACGUCC AD-20512 597 194GACGUGCAGCUGAUGAAGG 706 CCUUCAUCAGCUGCACGUC AD-20513 598 195ACGUGCAGCUGAUGAAGGG 707 CCCUUCAUCAGCUGCACGU AD-20514 660 196GAGAAUGAGGCUCUGUGGC 708 GCCACAGAGCCUCAUUCUC AD-20515 661 197AGAAUGAGGCUCUGUGGCG 709 CGCCACAGAGCCUCAUUCU AD-20516 662 198GAAUGAGGCUCUGUGGCGG 710 CCGCCACAGAGCCUCAUUC AD-20517 663 199AAUGAGGCUCUGUGGCGGG 711 CCCGCCACAGAGCCUCAUU AD-20518 664 200AUGAGGCUCUGUGGCGGGA 712 UCCCGCCACAGAGCCUCAU AD-20519 665 201UGAGGCUCUGUGGCGGGAG 713 CUCCCGCCACAGAGCCUCA AD-20520 666 202GAGGCUCUGUGGCGGGAGG 714 CCUCCCGCCACAGAGCCUC AD-20521 667 203AGGCUCUGUGGCGGGAGGU 715 ACCUCCCGCCACAGAGCCU AD-20522 668 204GGCUCUGUGGCGGGAGGUG 716 CACCUCCCGCCACAGAGCC AD-20523 669 205GCUCUGUGGCGGGAGGUGG 717 CCACCUCCCGCCACAGAGC AD-20524 670 206CUCUGUGGCGGGAGGUGGC 718 GCCACCUCCCGCCACAGAG AD-20525 671 207UCUGUGGCGGGAGGUGGCC 719 GGCCACCUCCCGCCACAGA AD-20526 672 208CUGUGGCGGGAGGUGGCCA 720 UGGCCACCUCCCGCCACAG AD-20527 673 209UGUGGCGGGAGGUGGCCAG 721 CUGGCCACCUCCCGCCACA AD-20528 674 210GUGGCGGGAGGUGGCCAGC 722 GCUGGCCACCUCCCGCCAC AD-20529 675 211UGGCGGGAGGUGGCCAGCC 723 GGCUGGCCACCUCCCGCCA AD-20530 676 212GGCGGGAGGUGGCCAGCCU 724 AGGCUGGCCACCUCCCGCC AD-20531 677 213GCGGGAGGUGGCCAGCCUU 725 AAGGCUGGCCACCUCCCGC AD-20532 678 214CGGGAGGUGGCCAGCCUUC 726 GAAGGCUGGCCACCUCCCG AD-20533 679 215GGGAGGUGGCCAGCCUUCG 727 CGAAGGCUGGCCACCUCCC AD-20534 680 216GGAGGUGGCCAGCCUUCGG 728 CCGAAGGCUGGCCACCUCC AD-20535 681 217GAGGUGGCCAGCCUUCGGC 729 GCCGAAGGCUGGCCACCUC AD-20536 682 218AGGUGGCCAGCCUUCGGCA 730 UGCCGAAGGCUGGCCACCU AD-20537 683 219GGUGGCCAGCCUUCGGCAG 731 CUGCCGAAGGCUGGCCACC AD-20538 684 220GUGGCCAGCCUUCGGCAGA 732 UCUGCCGAAGGCUGGCCAC AD-20539 685 221UGGCCAGCCUUCGGCAGAA 733 UUCUGCCGAAGGCUGGCCA R0035 686 222GGCCAGCCUUCGGCAGAAG 734 CUUCUGCCGAAGGCUGGCC R0036 687 223GCCAGCCUUCGGCAGAAGC 735 GCUUCUGCCGAAGGCUGGC R0037 688 224CCAGCCUUCGGCAGAAGCA 736 UGCUUCUGCCGAAGGCUGG R0038 689 225CAGCCUUCGGCAGAAGCAU 737 AUGCUUCUGCCGAAGGCUG AD-20540 690 226AGCCUUCGGCAGAAGCAUG 738 CAUGCUUCUGCCGAAGGCU AD-20541 691 227GCCUUCGGCAGAAGCAUGC 739 GCAUGCUUCUGCCGAAGGC AD-20542 692 228CCUUCGGCAGAAGCAUGCC 740 GGCAUGCUUCUGCCGAAGG AD-20543 693 229CUUCGGCAGAAGCAUGCCC 741 GGGCAUGCUUCUGCCGAAG AD-20544 694 230UUCGGCAGAAGCAUGCCCA 742 UGGGCAUGCUUCUGCCGAA AD-20545 695 231UCGGCAGAAGCAUGCCCAG 743 CUGGGCAUGCUUCUGCCGA AD-20546 696 232CGGCAGAAGCAUGCCCAGC 744 GCUGGGCAUGCUUCUGCCG AD-20547 697 233GGCAGAAGCAUGCCCAGCA 745 UGCUGGGCAUGCUUCUGCC AD-20548 698 234GCAGAAGCAUGCCCAGCAA 746 UUGCUGGGCAUGCUUCUGC AD-20549 699 235CAGAAGCAUGCCCAGCAAC 747 GUUGCUGGGCAUGCUUCUG AD-20550 700 236AGAAGCAUGCCCAGCAACA 748 UGUUGCUGGGCAUGCUUCU AD-20551 701 237GAAGCAUGCCCAGCAACAG 749 CUGUUGCUGGGCAUGCUUC AD-20552 702 238AAGCAUGCCCAGCAACAGA 750 UCUGUUGCUGGGCAUGCUU AD-20553 703 239AGCAUGCCCAGCAACAGAA 751 UUCUGUUGCUGGGCAUGCU AD-20554 704 240GCAUGCCCAGCAACAGAAA 752 UUUCUGUUGCUGGGCAUGC AD-20555 705 241CAUGCCCAGCAACAGAAAG 753 CUUUCUGUUGCUGGGCAUG AD-20556 706 242AUGCCCAGCAACAGAAAGU 754 ACUUUCUGUUGCUGGGCAU AD-20557 707 243UGCCCAGCAACAGAAAGUC 755 GACUUUCUGUUGCUGGGCA R0039 708 244GCCCAGCAACAGAAAGUCG 756 CGACUUUCUGUUGCUGGGC R0040 709 245CCCAGCAACAGAAAGUCGU 757 ACGACUUUCUGUUGCUGGG R0041 710 246CCAGCAACAGAAAGUCGUC 758 GACGACUUUCUGUUGCUGG R0042 711 247CAGCAACAGAAAGUCGUCA 759 UGACGACUUUCUGUUGCUG R0043 712 248AGCAACAGAAAGUCGUCAA 760 UUGACGACUUUCUGUUGCU R0044 713 249GCAACAGAAAGUCGUCAAC 761 GUUGACGACUUUCUGUUGC R0045 714 250CAACAGAAAGUCGUCAACA 762 UGUUGACGACUUUCUGUUG R0046 715 251AACAGAAAGUCGUCAACAA 763 UUGUUGACGACUUUCUGUU R0047 716 252ACAGAAAGUCGUCAACAAG 764 CUUGUUGACGACUUUCUGU R0048 717 253CAGAAAGUCGUCAACAAGC 765 GCUUGUUGACGACUUUCUG R0049 718 254AGAAAGUCGUCAACAAGCU 766 AGCUUGUUGACGACUUUCU R0050 719 255GAAAGUCGUCAACAAGCUC 767 GAGCUUGUUGACGACUUUC R0051 720 256AAAGUCGUCAACAAGCUCA 768 UGAGCUUGUUGACGACUUU R0052 721 257AAGUCGUCAACAAGCUCAU 769 AUGAGCUUGUUGACGACUU R0053 722 258AGUCGUCAACAAGCUCAUU 770 AAUGAGCUUGUUGACGACU R0054 723 259GUCGUCAACAAGCUCAUUC 771 GAAUGAGCUUGUUGACGAC R0055 724 260UCGUCAACAAGCUCAUUCA 772 UGAAUGAGCUUGUUGACGA R0056 725 261CGUCAACAAGCUCAUUCAG 773 CUGAAUGAGCUUGUUGACG R0057 726 262GUCAACAAGCUCAUUCAGU 774 ACUGAAUGAGCUUGUUGAC R0058 727 263UCAACAAGCUCAUUCAGUU 775 AACUGAAUGAGCUUGUUGA R0059 728 264CAACAAGCUCAUUCAGUUC 776 GAACUGAAUGAGCUUGUUG R0060 729 265AACAAGCUCAUUCAGUUCC 777 GGAACUGAAUGAGCUUGUU R0061 730 266ACAAGCUCAUUCAGUUCCU 778 AGGAACUGAAUGAGCUUGU AD-20558 731 267CAAGCUCAUUCAGUUCCUG 779 CAGGAACUGAAUGAGCUUG AD-20559 732 268AAGCUCAUUCAGUUCCUGA 780 UCAGGAACUGAAUGAGCUU AD-20560 733 269AGCUCAUUCAGUUCCUGAU 781 AUCAGGAACUGAAUGAGCU AD-20561 734 270GCUCAUUCAGUUCCUGAUC 782 GAUCAGGAACUGAAUGAGC AD-20562 735 271CUCAUUCAGUUCCUGAUCU 783 AGAUCAGGAACUGAAUGAG AD-20563 736 272UCAUUCAGUUCCUGAUCUC 784 GAGAUCAGGAACUGAAUGA AD-20564 737 273CAUUCAGUUCCUGAUCUCA 785 UGAGAUCAGGAACUGAAUG AD-20565 738 274AUUCAGUUCCUGAUCUCAC 786 GUGAGAUCAGGAACUGAAU AD-20566 739 275UUCAGUUCCUGAUCUCACU 787 AGUGAGAUCAGGAACUGAA AD-20567 740 276UCAGUUCCUGAUCUCACUG 788 CAGUGAGAUCAGGAACUGA AD-20568 741 277CAGUUCCUGAUCUCACUGG 789 CCAGUGAGAUCAGGAACUG AD-20569 742 278AGUUCCUGAUCUCACUGGU 790 ACCAGUGAGAUCAGGAACU AD-20571 744 279UUCCUGAUCUCACUGGUGC 791 GCACCAGUGAGAUCAGGAA AD-20572 745 280UCCUGAUCUCACUGGUGCA 792 UGCACCAGUGAGAUCAGGA AD-20573 746 281CCUGAUCUCACUGGUGCAG 793 CUGCACCAGUGAGAUCAGG AD-20574 747 2044CUGAUCUCACUGGUGCAGU 2045 ACUGCACCAGUGAGAUCAG AD-20575 748 282UGAUCUCACUGGUGCAGUC 794 GACUGCACCAGUGAGAUCA AD-20576 749 283GAUCUCACUGGUGCAGUCA 795 UGACUGCACCAGUGAGAUC AD-20577 750 284AUCUCACUGGUGCAGUCAA 796 UUGACUGCACCAGUGAGAU AD-20578 751 285UCUCACUGGUGCAGUCAAA 797 UUUGACUGCACCAGUGAGA AD-20579 752 286CUCACUGGUGCAGUCAAAC 798 GUUUGACUGCACCAGUGAG AD-20581 754 287CACUGGUGCAGUCAAACCG 799 CGGUUUGACUGCACCAGUG AD-20582 755 288ACUGGUGCAGUCAAACCGG 800 CCGGUUUGACUGCACCAGU AD-20625 756 289CUGGUGCAGUCAAACCGGA 801 UCCGGUUUGACUGCACCAG AD-20626 757 290UGGUGCAGUCAAACCGGAU 802 AUCCGGUUUGACUGCACCA AD-20627 758 291GGUGCAGUCAAACCGGAUC 803 GAUCCGGUUUGACUGCACC AD-20628 759 292GUGCAGUCAAACCGGAUCC 804 GGAUCCGGUUUGACUGCAC AD-20629 760 293UGCAGUCAAACCGGAUCCU 805 AGGAUCCGGUUUGACUGCA AD-20630 761 294GCAGUCAAACCGGAUCCUG 806 CAGGAUCCGGUUUGACUGC AD-20631 762 295CAGUCAAACCGGAUCCUGG 807 CCAGGAUCCGGUUUGACUG AD-20632 763 296AGUCAAACCGGAUCCUGGG 808 CCCAGGAUCCGGUUUGACU AD-20633 781 297GGGUGAAGAGAAAGAUCCC 809 GGGAUCUUUCUCUUCACCC AD-20634 799 298CCCUGAUGCUGAACGACAG 810 CUGUCGUUCAGCAUCAGGG AD-20635 800 299CCUGAUGCUGAACGACAGU 811 ACUGUCGUUCAGCAUCAGG AD-20636 801 300CUGAUGCUGAACGACAGUG 812 CACUGUCGUUCAGCAUCAG AD-20637 802 301UGAUGCUGAACGACAGUGG 813 CCACUGUCGUUCAGCAUCA AD-20638 803 302GAUGCUGAACGACAGUGGC 814 GCCACUGUCGUUCAGCAUC AD-20639 804 303AUGCUGAACGACAGUGGCU 815 AGCCACUGUCGUUCAGCAU AD-20640 805 304UGCUGAACGACAGUGGCUC 816 GAGCCACUGUCGUUCAGCA AD-20641 806 305GCUGAACGACAGUGGCUCA 817 UGAGCCACUGUCGUUCAGC AD-20642 807 306CUGAACGACAGUGGCUCAG 818 CUGAGCCACUGUCGUUCAG AD-20643 808 307UGAACGACAGUGGCUCAGC 819 GCUGAGCCACUGUCGUUCA AD-20644 809 308GAACGACAGUGGCUCAGCA 820 UGCUGAGCCACUGUCGUUC AD-20645 810 309AACGACAGUGGCUCAGCAC 821 GUGCUGAGCCACUGUCGUU AD-20646 811 310ACGACAGUGGCUCAGCACA 822 UGUGCUGAGCCACUGUCGU AD-20647 812 311CGACAGUGGCUCAGCACAU 823 AUGUGCUGAGCCACUGUCG AD-20648 813 312GACAGUGGCUCAGCACAUU 824 AAUGUGCUGAGCCACUGUC AD-20649 814 313ACAGUGGCUCAGCACAUUC 825 GAAUGUGCUGAGCCACUGU AD-20650 815 314CAGUGGCUCAGCACAUUCC 826 GGAAUGUGCUGAGCCACUG AD-20651 816 315AGUGGCUCAGCACAUUCCA 827 UGGAAUGUGCUGAGCCACU AD-20652 817 316GUGGCUCAGCACAUUCCAU 828 AUGGAAUGUGCUGAGCCAC AD-20653 818 317UGGCUCAGCACAUUCCAUG 829 CAUGGAAUGUGCUGAGCCA AD-20654 819 318GGCUCAGCACAUUCCAUGC 830 GCAUGGAAUGUGCUGAGCC AD-20655 820 319GCUCAGCACAUUCCAUGCC 831 GGCAUGGAAUGUGCUGAGC AD-20656 821 320CUCAGCACAUUCCAUGCCC 832 GGGCAUGGAAUGUGCUGAG AD-20657 822 321UCAGCACAUUCCAUGCCCA 833 UGGGCAUGGAAUGUGCUGA AD-20658 823 322CAGCACAUUCCAUGCCCAA 834 UUGGGCAUGGAAUGUGCUG AD-20659 824 323AGCACAUUCCAUGCCCAAG 835 CUUGGGCAUGGAAUGUGCU AD-20660 825 324GCACAUUCCAUGCCCAAGU 836 ACUUGGGCAUGGAAUGUGC AD-20661 826 325CACAUUCCAUGCCCAAGUA 837 UACUUGGGCAUGGAAUGUG AD-20284 827 326ACAUUCCAUGCCCAAGUAU 838 AUACUUGGGCAUGGAAUGU AD-20662 847 327GCCGGCAGUUCUCCCUGGA 839 UCCAGGGAGAACUGCCGGC AD-20868 848 328CCGGCAGUUCUCCCUGGAG 840 CUCCAGGGAGAACUGCCGG AD-20663 849 329CGGCAGUUCUCCCUGGAGC 841 GCUCCAGGGAGAACUGCCG AD-20664 850 330GGCAGUUCUCCCUGGAGCA 842 UGCUCCAGGGAGAACUGCC AD-20665 851 331GCAGUUCUCCCUGGAGCAC 843 GUGCUCCAGGGAGAACUGC AD-20666 852 332CAGUUCUCCCUGGAGCACG 844 CGUGCUCCAGGGAGAACUG AD-20667 853 333AGUUCUCCCUGGAGCACGU 845 ACGUGCUCCAGGGAGAACU AD-20668 854 334GUUCUCCCUGGAGCACGUC 846 GACGUGCUCCAGGGAGAAC AD-20669 855 335UUCUCCCUGGAGCACGUCC 847 GGACGUGCUCCAGGGAGAA AD-20670 856 336UCUCCCUGGAGCACGUCCA 848 UGGACGUGCUCCAGGGAGA AD-20671 857 337CUCCCUGGAGCACGUCCAC 849 GUGGACGUGCUCCAGGGAG AD-20672 858 338UCCCUGGAGCACGUCCACG 850 CGUGGACGUGCUCCAGGGA AD-20673 859 339CCCUGGAGCACGUCCACGG 851 CCGUGGACGUGCUCCAGGG AD-20674 860 340CCUGGAGCACGUCCACGGC 852 GCCGUGGACGUGCUCCAGG AD-20675 861 341CUGGAGCACGUCCACGGCU 853 AGCCGUGGACGUGCUCCAG AD-20676 862 342UGGAGCACGUCCACGGCUC 854 GAGCCGUGGACGUGCUCCA R0062 863 343GGAGCACGUCCACGGCUCG 855 CGAGCCGUGGACGUGCUCC R0063 864 344GAGCACGUCCACGGCUCGG 856 CCGAGCCGUGGACGUGCUC R0064 865 345AGCACGUCCACGGCUCGGG 857 CCCGAGCCGUGGACGUGCU R0065 866 346GCACGUCCACGGCUCGGGC 858 GCCCGAGCCGUGGACGUGC R0066 867 347CACGUCCACGGCUCGGGCC 859 GGCCCGAGCCGUGGACGUG R0067 868 348ACGUCCACGGCUCGGGCCC 860 GGGCCCGAGCCGUGGACGU AD-20677 915 349AGCUCCAGCCUCUACGCCC 861 GGGCGUAGAGGCUGGAGCU R0068 954 350GGACCCAUCAUCUCCGACA 862 UGUCGGAGAUGAUGGGUCC R0069 955 351GACCCAUCAUCUCCGACAU 863 AUGUCGGAGAUGAUGGGUC R0070 956 352ACCCAUCAUCUCCGACAUC 864 GAUGUCGGAGAUGAUGGGU R0071 957 353CCCAUCAUCUCCGACAUCA 865 UGAUGUCGGAGAUGAUGGG R0072 958 354CCAUCAUCUCCGACAUCAC 866 GUGAUGUCGGAGAUGAUGG R0073 959 355CAUCAUCUCCGACAUCACC 867 GGUGAUGUCGGAGAUGAUG R0074 960 356AUCAUCUCCGACAUCACCG 868 CGGUGAUGUCGGAGAUGAU R0075 961 357UCAUCUCCGACAUCACCGA 869 UCGGUGAUGUCGGAGAUGA R0076 962 358CAUCUCCGACAUCACCGAG 870 CUCGGUGAUGUCGGAGAUG R0077 963 359AUCUCCGACAUCACCGAGC 871 GCUCGGUGAUGUCGGAGAU R0078 964 360UCUCCGACAUCACCGAGCU 872 AGCUCGGUGAUGUCGGAGA AD-20678 965 361CUCCGACAUCACCGAGCUG 873 CAGCUCGGUGAUGUCGGAG AD-20679 966 362UCCGACAUCACCGAGCUGG 874 CCAGCUCGGUGAUGUCGGA AD-20680 967 363CCGACAUCACCGAGCUGGC 875 GCCAGCUCGGUGAUGUCGG AD-20681 968 364CGACAUCACCGAGCUGGCU 876 AGCCAGCUCGGUGAUGUCG AD-20682 969 365GACAUCACCGAGCUGGCUC 877 GAGCCAGCUCGGUGAUGUC AD-20683 970 366ACAUCACCGAGCUGGCUCC 878 GGAGCCAGCUCGGUGAUGU AD-20684 971 367CAUCACCGAGCUGGCUCCU 879 AGGAGCCAGCUCGGUGAUG AD-20685 972 368AUCACCGAGCUGGCUCCUG 880 CAGGAGCCAGCUCGGUGAU AD-20686 973 369UCACCGAGCUGGCUCCUGC 881 GCAGGAGCCAGCUCGGUGA AD-20687 974 370CACCGAGCUGGCUCCUGCC 882 GGCAGGAGCCAGCUCGGUG AD-20688 975 371ACCGAGCUGGCUCCUGCCA 883 UGGCAGGAGCCAGCUCGGU AD-20689 976 372CCGAGCUGGCUCCUGCCAG 884 CUGGCAGGAGCCAGCUCGG AD-20690 977 373CGAGCUGGCUCCUGCCAGC 885 GCUGGCAGGAGCCAGCUCG AD-20691 978 374GAGCUGGCUCCUGCCAGCC 886 GGCUGGCAGGAGCCAGCUC AD-20692 979 375AGCUGGCUCCUGCCAGCCC 887 GGGCUGGCAGGAGCCAGCU AD-20693 1011 376GGCGGGAGCAUAGACGAGA 888 UCUCGUCUAUGCUCCCGCC AD-20694 1012 377GCGGGAGCAUAGACGAGAG 889 CUCUCGUCUAUGCUCCCGC AD-20695 1013 378CGGGAGCAUAGACGAGAGG 890 CCUCUCGUCUAUGCUCCCG AD-20696 1014 379GGGAGCAUAGACGAGAGGC 891 GCCUCUCGUCUAUGCUCCC AD-20697 1015 380GGAGCAUAGACGAGAGGCC 892 GGCCUCUCGUCUAUGCUCC AD-20698 1016 381GAGCAUAGACGAGAGGCCC 893 GGGCCUCUCGUCUAUGCUC AD-20699 1048 382CCCUGGUGCGUGUCAAGGA 894 UCCUUGACACGCACCAGGG AD-20700 1049 383CCUGGUGCGUGUCAAGGAG 895 CUCCUUGACACGCACCAGG AD-20701 1050 384CUGGUGCGUGUCAAGGAGG 896 CCUCCUUGACACGCACCAG AD-20702 1051 385UGGUGCGUGUCAAGGAGGA 897 UCCUCCUUGACACGCACCA AD-20869 1052 386GGUGCGUGUCAAGGAGGAG 898 CUCCUCCUUGACACGCACC AD-20703 1053 387GUGCGUGUCAAGGAGGAGC 899 GCUCCUCCUUGACACGCAC AD-20704 1054 388UGCGUGUCAAGGAGGAGCC 900 GGCUCCUCCUUGACACGCA AD-20705 1055 389GCGUGUCAAGGAGGAGCCC 901 GGGCUCCUCCUUGACACGC R0079 1074 390CCCAGCCCGCCUCAGAGCC 902 GGCUCUGAGGCGGGCUGGG R0080 1075 391CCAGCCCGCCUCAGAGCCC 903 GGGCUCUGAGGCGGGCUGG AD-20706 1329 392CACUUGGAUGCUAUGGACU 904 AGUCCAUAGCAUCCAAGUG AD-20707 1330 393ACUUGGAUGCUAUGGACUC 905 GAGUCCAUAGCAUCCAAGU AD-20709 1332 394UUGGAUGCUAUGGACUCCA 906 UGGAGUCCAUAGCAUCCAA AD-20710 1333 395UGGAUGCUAUGGACUCCAA 907 UUGGAGUCCAUAGCAUCCA AD-20711 1334 396GGAUGCUAUGGACUCCAAC 908 GUUGGAGUCCAUAGCAUCC AD-20712 1335 397GAUGCUAUGGACUCCAACC 909 GGUUGGAGUCCAUAGCAUC AD-20713 1336 398AUGCUAUGGACUCCAACCU 910 AGGUUGGAGUCCAUAGCAU AD-20714 1337 399UGCUAUGGACUCCAACCUG 911 CAGGUUGGAGUCCAUAGCA AD-20715 1338 400GCUAUGGACUCCAACCUGG 912 CCAGGUUGGAGUCCAUAGC AD-20716 1339 401CUAUGGACUCCAACCUGGA 913 UCCAGGUUGGAGUCCAUAG AD-20717 1359 402AACCUGCAGACCAUGCUGA 914 UCAGCAUGGUCUGCAGGUU AD-20718 1360 403ACCUGCAGACCAUGCUGAG 915 CUCAGCAUGGUCUGCAGGU AD-20719 1361 404CCUGCAGACCAUGCUGAGC 916 GCUCAGCAUGGUCUGCAGG AD-20720 1362 405CUGCAGACCAUGCUGAGCA 917 UGCUCAGCAUGGUCUGCAG AD-20721 1363 406UGCAGACCAUGCUGAGCAG 918 CUGCUCAGCAUGGUCUGCA AD-20722 1364 407GCAGACCAUGCUGAGCAGC 919 GCUGCUCAGCAUGGUCUGC AD-20723 1365 408CAGACCAUGCUGAGCAGCC 920 GGCUGCUCAGCAUGGUCUG AD-20724 1366 409AGACCAUGCUGAGCAGCCA 921 UGGCUGCUCAGCAUGGUCU AD-20725 1367 410GACCAUGCUGAGCAGCCAC 922 GUGGCUGCUCAGCAUGGUC AD-20726 1368 411ACCAUGCUGAGCAGCCACG 923 CGUGGCUGCUCAGCAUGGU AD-20727 1369 412CCAUGCUGAGCAGCCACGG 924 CCGUGGCUGCUCAGCAUGG AD-20728 1370 413CAUGCUGAGCAGCCACGGC 925 GCCGUGGCUGCUCAGCAUG AD-20729 1371 414AUGCUGAGCAGCCACGGCU 926 AGCCGUGGCUGCUCAGCAU AD-20730 1372 415UGCUGAGCAGCCACGGCUU 927 AAGCCGUGGCUGCUCAGCA AD-20731 1373 416GCUGAGCAGCCACGGCUUC 928 GAAGCCGUGGCUGCUCAGC AD-20732 1374 417CUGAGCAGCCACGGCUUCA 929 UGAAGCCGUGGCUGCUCAG AD-20733 1375 418UGAGCAGCCACGGCUUCAG 930 CUGAAGCCGUGGCUGCUCA R0081 1376 419GAGCAGCCACGGCUUCAGC 931 GCUGAAGCCGUGGCUGCUC R0082 1377 420AGCAGCCACGGCUUCAGCG 932 CGCUGAAGCCGUGGCUGCU R0083 1378 421GCAGCCACGGCUUCAGCGU 933 ACGCUGAAGCCGUGGCUGC AD-20734 1379 422CAGCCACGGCUUCAGCGUG 934 CACGCUGAAGCCGUGGCUG AD-20735 1380 423AGCCACGGCUUCAGCGUGG 935 CCACGCUGAAGCCGUGGCU AD-20736 1381 424GCCACGGCUUCAGCGUGGA 936 UCCACGCUGAAGCCGUGGC AD-20737 1382 425CCACGGCUUCAGCGUGGAC 937 GUCCACGCUGAAGCCGUGG AD-20738 1383 426CACGGCUUCAGCGUGGACA 938 UGUCCACGCUGAAGCCGUG AD-20739 1384 427ACGGCUUCAGCGUGGACAC 939 GUGUCCACGCUGAAGCCGU AD-20740 1385 428CGGCUUCAGCGUGGACACC 940 GGUGUCCACGCUGAAGCCG AD-20741 1386 429GGCUUCAGCGUGGACACCA 941 UGGUGUCCACGCUGAAGCC AD-20742 1387 430GCUUCAGCGUGGACACCAG 942 CUGGUGUCCACGCUGAAGC AD-20743 1407 431GCCCUGCUGGACCUGUUCA 943 UGAACAGGUCCAGCAGGGC AD-20744 1408 432CCCUGCUGGACCUGUUCAG 944 CUGAACAGGUCCAGCAGGG AD-20745 1409 433CCUGCUGGACCUGUUCAGC 945 GCUGAACAGGUCCAGCAGG AD-20746 1410 434CUGCUGGACCUGUUCAGCC 946 GGCUGAACAGGUCCAGCAG AD-20747 1411 435UGCUGGACCUGUUCAGCCC 947 GGGCUGAACAGGUCCAGCA AD-20748 1428 436CCCUCGGUGACCGUGCCCG 948 CGGGCACGGUCACCGAGGG AD-20749 1429 437CCUCGGUGACCGUGCCCGA 949 UCGGGCACGGUCACCGAGG AD-20750 1430 438CUCGGUGACCGUGCCCGAC 950 GUCGGGCACGGUCACCGAG AD-20751 1431 439UCGGUGACCGUGCCCGACA 951 UGUCGGGCACGGUCACCGA AD-20752 1432 440CGGUGACCGUGCCCGACAU 952 AUGUCGGGCACGGUCACCG AD-20753 1433 441GGUGACCGUGCCCGACAUG 953 CAUGUCGGGCACGGUCACC AD-20754 1434 442GUGACCGUGCCCGACAUGA 954 UCAUGUCGGGCACGGUCAC AD-20755 1435 443UGACCGUGCCCGACAUGAG 955 CUCAUGUCGGGCACGGUCA AD-20756 1436 444GACCGUGCCCGACAUGAGC 956 GCUCAUGUCGGGCACGGUC AD-20757 1437 445ACCGUGCCCGACAUGAGCC 957 GGCUCAUGUCGGGCACGGU AD-20758 1438 446CCGUGCCCGACAUGAGCCU 958 AGGCUCAUGUCGGGCACGG AD-20759 1439 447CGUGCCCGACAUGAGCCUG 959 CAGGCUCAUGUCGGGCACG AD-20760 1440 448GUGCCCGACAUGAGCCUGC 960 GCAGGCUCAUGUCGGGCAC AD-20761 1441 449UGCCCGACAUGAGCCUGCC 961 GGCAGGCUCAUGUCGGGCA AD-20762 1442 450GCCCGACAUGAGCCUGCCU 962 AGGCAGGCUCAUGUCGGGC AD-20763 1443 451CCCGACAUGAGCCUGCCUG 963 CAGGCAGGCUCAUGUCGGG AD-20764 1444 452CCGACAUGAGCCUGCCUGA 964 UCAGGCAGGCUCAUGUCGG AD-20765 1445 453CGACAUGAGCCUGCCUGAC 965 GUCAGGCAGGCUCAUGUCG AD-20766 1446 454GACAUGAGCCUGCCUGACC 966 GGUCAGGCAGGCUCAUGUC AD-20767 1447 455ACAUGAGCCUGCCUGACCU 967 AGGUCAGGCAGGCUCAUGU AD-20768 1448 456CAUGAGCCUGCCUGACCUU 968 AAGGUCAGGCAGGCUCAUG AD-20769 1449 457AUGAGCCUGCCUGACCUUG 969 CAAGGUCAGGCAGGCUCAU AD-20770 1450 458UGAGCCUGCCUGACCUUGA 970 UCAAGGUCAGGCAGGCUCA AD-20771 1451 459GAGCCUGCCUGACCUUGAC 971 GUCAAGGUCAGGCAGGCUC AD-20772 1452 460AGCCUGCCUGACCUUGACA 972 UGUCAAGGUCAGGCAGGCU AD-20773 1453 461GCCUGCCUGACCUUGACAG 973 CUGUCAAGGUCAGGCAGGC AD-20774 1454 462CCUGCCUGACCUUGACAGC 974 GCUGUCAAGGUCAGGCAGG AD-20775 1455 463CUGCCUGACCUUGACAGCA 975 UGCUGUCAAGGUCAGGCAG AD-20776 1456 464UGCCUGACCUUGACAGCAG 976 CUGCUGUCAAGGUCAGGCA AD-20777 1457 465GCCUGACCUUGACAGCAGC 977 GCUGCUGUCAAGGUCAGGC AD-20778 1458 466CCUGACCUUGACAGCAGCC 978 GGCUGCUGUCAAGGUCAGG AD-20779 1459 467CUGACCUUGACAGCAGCCU 979 AGGCUGCUGUCAAGGUCAG AD-20780 1460 468UGACCUUGACAGCAGCCUG 980 CAGGCUGCUGUCAAGGUCA AD-20781 1461 469GACCUUGACAGCAGCCUGG 981 CCAGGCUGCUGUCAAGGUC AD-20782 1462 470ACCUUGACAGCAGCCUGGC 982 GCCAGGCUGCUGUCAAGGU AD-20783 1482 471AGUAUCCAAGAGCUCCUGU 983 ACAGGAGCUCUUGGAUACU AD-20784 1483 472GUAUCCAAGAGCUCCUGUC 984 GACAGGAGCUCUUGGAUAC AD-20785 1484 473UAUCCAAGAGCUCCUGUCU 985 AGACAGGAGCUCUUGGAUA AD-20786 1485 474AUCCAAGAGCUCCUGUCUC 986 GAGACAGGAGCUCUUGGAU AD-20787 1486 475UCCAAGAGCUCCUGUCUCC 987 GGAGACAGGAGCUCUUGGA AD-20788 1487 476CCAAGAGCUCCUGUCUCCC 988 GGGAGACAGGAGCUCUUGG R0084 1533 477GAGAACAGCAGCCCGGAUU 989 AAUCCGGGCUGCUGUUCUC R0085 1534 478AGAACAGCAGCCCGGAUUC 990 GAAUCCGGGCUGCUGUUCU R0086 1535 479GAACAGCAGCCCGGAUUCA 991 UGAAUCCGGGCUGCUGUUC R0087 1536 480AACAGCAGCCCGGAUUCAG 992 CUGAAUCCGGGCUGCUGUU R0088 1537 481ACAGCAGCCCGGAUUCAGG 993 CCUGAAUCCGGGCUGCUGU R0089 1538 482CAGCAGCCCGGAUUCAGGG 994 CCCUGAAUCCGGGCUGCUG R0090 1539 483AGCAGCCCGGAUUCAGGGA 995 UCCCUGAAUCCGGGCUGCU R0091 1540 484GCAGCCCGGAUUCAGGGAA 996 UUCCCUGAAUCCGGGCUGC R0092 1541 485CAGCCCGGAUUCAGGGAAG 997 CUUCCCUGAAUCCGGGCUG R0093 1542 486AGCCCGGAUUCAGGGAAGC 998 GCUUCCCUGAAUCCGGGCU R0094 1543 487GCCCGGAUUCAGGGAAGCA 999 UGCUUCCCUGAAUCCGGGC R0095 1544 488CCCGGAUUCAGGGAAGCAG 1000 CUGCUUCCCUGAAUCCGGG R0096 1545 489CCGGAUUCAGGGAAGCAGC 1001 GCUGCUUCCCUGAAUCCGG R0097 1546 490CGGAUUCAGGGAAGCAGCU 1002 AGCUGCUUCCCUGAAUCCG AD-20789 1547 491GGAUUCAGGGAAGCAGCUG 1003 CAGCUGCUUCCCUGAAUCC AD-20790 1548 492GAUUCAGGGAAGCAGCUGG 1004 CCAGCUGCUUCCCUGAAUC AD-20791 1549 493AUUCAGGGAAGCAGCUGGU 1005 ACCAGCUGCUUCCCUGAAU AD-20792 1602 494CCCGGCUCCGUGGACACCG 1006 CGGUGUCCACGGAGCCGGG AD-20793 1603 495CCGGCUCCGUGGACACCGG 1007 CCGGUGUCCACGGAGCCGG AD-20794 1604 496CGGCUCCGUGGACACCGGG 1008 CCCGGUGUCCACGGAGCCG AD-20795 1605 497GGCUCCGUGGACACCGGGA 1009 UCCCGGUGUCCACGGAGCC AD-20796 1606 498GCUCCGUGGACACCGGGAG 1010 CUCCCGGUGUCCACGGAGC AD-20870 1607 499CUCCGUGGACACCGGGAGC 1011 GCUCCCGGUGUCCACGGAG AD-20871 1608 500UCCGUGGACACCGGGAGCA 1012 UGCUCCCGGUGUCCACGGA AD-20872 1633 501UGCCGGUGCUGUUUGAGCU 1013 AGCUCAAACAGCACCGGCA AD-20797 1634 502GCCGGUGCUGUUUGAGCUG 1014 CAGCUCAAACAGCACCGGC AD-20798 1635 503CCGGUGCUGUUUGAGCUGG 1015 CCAGCUCAAACAGCACCGG AD-20799 1636 504CGGUGCUGUUUGAGCUGGG 1016 CCCAGCUCAAACAGCACCG AD-20873 1698 505CCCACCAUCUCCCUGCUGA 1017 UCAGCAGGGAGAUGGUGGG AD-20800 1699 506CCACCAUCUCCCUGCUGAC 1018 GUCAGCAGGGAGAUGGUGG AD-20801 1700 507CACCAUCUCCCUGCUGACA 1019 UGUCAGCAGGGAGAUGGUG R0098 1725 508GAGCCUCCCAAAGCCAAGG 1020 CCUUGGCUUUGGGAGGCUC R0099 1726 509AGCCUCCCAAAGCCAAGGA 1021 UCCUUGGCUUUGGGAGGCU R0100 1727 510GCCUCCCAAAGCCAAGGAC 1022 GUCCUUGGCUUUGGGAGGC R0101 1728 511CCUCCCAAAGCCAAGGACC 1023 GGUCCUUGGCUUUGGGAGG R0102 1729 512CUCCCAAAGCCAAGGACCC 1024 GGGUCCUUGGCUUUGGGAG AD-20278 2053GCAGGUUGUUCAUAGUCAG 2064 CUGACUAUGAACAACCUGC AD-20279 2054CAGGUUGUUCAUAGUCAGA 2065 UCUGACUAUGAACAACCUG AD-20280 2055AGGUUGUUCAUAGUCAGAA 2066 UUCUGACUAUGAACAACCU AD-20281 2056GCCCAAGUACUUCAAGCAC 2067 GUGCUUGAAGUACUUGGGC AD-20282 2057CCCAAGUACUUCAAGCACA 2068 UGUGCUUGAAGUACUUGGG AD-20283 2058CCAAGUACUUCAAGCACAA 2069 UUGUGCUUGAAGUACUUGG AD-20377 2059CAACAACAUGGCCAGCUUC 2070 GAAGCUGGCCAUGUUGUUG AD-20570 2060GUUCCUGAUCUCACUGGUG 2071 CACCAGUGAGAUCAGGAAC AD-20580 2061UCACUGGUGCAGUCAAACC 2072 GGUUUGACUGCACCAGUGA AD-20597 2062UUGGUCAGGAAGGCCGGGA 2073 UCCCGGCCUUCCUGACCAA AD-20598 2063CCCGGCCUUCCUGACCAAG 2074 CUUGGUCAGGAAGGCCGGG

Modifications of the sequences in Table 1 are easily conceived by one ofskill in the art. Example and non-limiting modifications of thesesequences were conceived. These are listed in Tables 2 and 3. Additionalmodifications are contemplated.

For the modified sequences listed in Tables 2 and 3, some modificationswere placed at sites predicted to be sensitive to endonucleases. Somemodifications were designed to eliminate an immune response to the siRNAwhile preserving activity. In general, the sense strand was heavilymodified, and the antisense strand lightly modified. Some modificationsserve more than one purpose. Table 2 lists RNAi agents prepared withthese modified sequences.

Subsequent to screening, a full-length HSF1 gene sequence fromcynomolgus monkey was produced (see above), and used to assess identityof human/cyno 19-mer sequences. Of 406 19-mers screened for single doseefficacy in WI38 and Hela cells, 375 are perfect matches between humanand cyno (e.g., zero mismatches).

TABLE 2 HSF1 RNAi agents (with modified sequences) Duplex SEQ ID SEQ IDName Position NO Sense 5′-3′modified NO Antisense 5′-3′ modifiedAD-20594 209 1033 cAAcGucccGGccuuccuGdTdT 1545 cAGGAAGGCCGGGACGUUGdTdTAD-20595 210 1034 UcAGGAAGGCCGGGACGUUdTdT 1546 AAcGucccGGccuuccuGAdTdTAD-20596 211 1035 AcGucccGGccuuccuGAcdTdT 1547 GUcAGGAAGGCCGGGACGUdTdTAD-20285 212 1036 cGucccGGccuuccuGAccdTdT 1548 GGUcAGGAAGGCCGGGACGdTdTAD-20286 213 1037 GucccGGccuuccuGAccAdTdT 1549 UGGUcAGGAAGGCCGGGACdTdTAD-20287 216 1038 ccGGccuuccuGAccAAGcdTdT 1550 GCUUGGUcAGGAAGGCCGGdTdTAD-20288 217 1039 cGGccuuccuGAccAAGcudTdT 1551 AGCUUGGUcAGGAAGGCCGdTdTAD-20289 218 1040 GGccuuccuGAccAAGcuGdTdT 1552 cAGCUUGGUcAGGAAGGCCdTdTAD-20290 219 1041 GccuuccuGAccAAGcuGudTdT 1553 AcAGCUUGGUcAGGAAGGCdTdTAD-20291 220 1042 ccuuccuGAccAAGcuGuGdTdT 1554 cAcAGCUUGGUcAGGAAGGdTdTAD-20292 221 1043 cuuccuGAccAAGcuGuGGdTdT 1555 CcAcAGCUUGGUcAGGAAGdTdTAD-20293 222 1044 uuccuGAccAAGcuGuGGAdTdT 1556 UCcAcAGCUUGGUcAGGAAdTdTAD-20294 223 1045 uccuGAccAAGcuGuGGAcdTdT 1557 GUCcAcAGCUUGGUcAGGAdTdTAD-20295 224 1046 ccuGAccAAGcuGuGGAccdTdT 1558 GGUCcAcAGCUUGGUcAGGdTdTAD-20296 225 1047 cuGAccAAGcuGuGGAcccdTdT 1559 GGGUCcAcAGCUUGGUcAGdTdTAD-20297 226 1048 uGAccAAGcuGuGGAcccudTdT 1560 AGGGUCcAcAGCUUGGUcAdTdTAD-20298 227 1049 GAccAAGcuGuGGAcccucdTdT 1561 GAGGGUCcAcAGCUUGGUCdTdTAD-20299 228 1050 AccAAGcuGuGGAcccucGdTdT 1562 CGAGGGUCcAcAGCUUGGUdTdTAD-20300 229 1051 ccAAGcuGuGGAcccucGudTdT 1563 ACGAGGGUCcAcAGCUUGGdTdTAD-20301 230 1052 cAAGcuGuGGAcccucGuGdTdT 1564 cACGAGGGUCcAcAGCUUGdTdTAD-20302 231 1053 AAGcuGuGGAcccucGuGAdTdT 1565 UcACGAGGGUCcAcAGCUUdTdTAD-20303 232 1054 AGcuGuGGAcccucGuGAGdTdT 1566 CUcACGAGGGUCcAcAGCUdTdTAD-20304 233 1055 GcuGuGGAcccucGuGAGcdTdT 1567 GCUcACGAGGGUCcAcAGCdTdTAD-20305 234 1056 cuGuGGAcccucGuGAGcGdTdT 1568 CGCUcACGAGGGUCcAcAGdTdTAD-20306 235 1057 uGuGGAcccucGuGAGcGAdTdT 1569 UCGCUcACGAGGGUCcAcAdTdTAD-20307 236 1058 GuGGAcccucGuGAGcGAcdTdT 1570 GUCGCUcACGAGGGUCcACdTdTAD-20308 237 1059 uGGAcccucGuGAGcGAccdTdT 1571 GGUCGCUcACGAGGGUCcAdTdTAD-20309 238 1060 GGAcccucGuGAGcGAcccdTdT 1572 GGGUCGCUcACGAGGGUCCdTdTAD-20310 239 1061 GAcccucGuGAGcGAcccGdTdT 1573 CGGGUCGCUcACGAGGGUCdTdTAD-20311 240 1062 AcccucGuGAGcGAcccGGdTdT 1574 CCGGGUCGCUcACGAGGGUdTdTAD-20312 241 1063 cccucGuGAGcGAcccGGAdTdT 1575 UCCGGGUCGCUcACGAGGGdTdTAD-20313 242 1064 ccucGuGAGcGAcccGGAcdTdT 1576 GUCCGGGUCGCUcACGAGGdTdTAD-20314 243 1065 cucGuGAGcGAcccGGAcAdTdT 1577 UGUCCGGGUCGCUcACGAGdTdTAD-20315 244 1066 ucGuGAGcGAcccGGAcAcdTdT 1578 GUGUCCGGGUCGCUcACGAdTdTAD-20316 245 1067 cGuGAGcGAcccGGAcAccdTdT 1579 GGUGUCCGGGUCGCUcACGdTdTAD-20317 246 1068 GuGAGcGAcccGGAcAccGdTdT 1580 CGGUGUCCGGGUCGCUcACdTdTAD-20318 247 1069 uGAGcGAcccGGAcAccGAdTdT 1581 UCGGUGUCCGGGUCGCUcAdTdTAD-20319 270 1073 cucAucuGcuGGAGcccGAdTdT 1585 UCGGGCUCcAGcAGAUGAGdTdTAD-20320 271 1074 ucAucuGcuGGAGcccGAGdTdT 1586 CUCGGGCUCcAGcAGAUGAdTdTAD-20344 306 1075 GuGuucGAccAGGGccAGudTdT 1587 ACUGGCCCUGGUCGAAcACdTdTAD-20345 307 1076 uGuucGAccAGGGccAGuudTdT 1588 AACUGGCCCUGGUCGAAcAdTdTAD-20346 309 1078 uucGAccAGGGccAGuuuGdTdT 1590 cAAACUGGCCCUGGUCGAAdTdTAD-20347 310 1079 ucGAccAGGGccAGuuuGcdTdT 1591 GcAAACUGGCCCUGGUCGAdTdTAD-20348 311 1080 cGAccAGGGccAGuuuGccdTdT 1592 GGcAAACUGGCCCUGGUCGdTdTAD-20349 312 1081 GAccAGGGccAGuuuGccAdTdT 1593 UGGcAAACUGGCCCUGGUCdTdTAD-20350 313 1082 AccAGGGccAGuuuGccAAdTdT 1594 UUGGcAAACUGGCCCUGGUdTdTAD-20351 314 1083 ccAGGGccAGuuuGccAAGdTdT 1595 CUUGGcAAACUGGCCCUGGdTdTAD-20352 315 1084 cAGGGccAGuuuGccAAGGdTdT 1596 CCUUGGcAAACUGGCCCUGdTdTAD-20353 316 1085 AGGGccAGuuuGccAAGGAdTdT 1597 UCCUUGGcAAACUGGCCCUdTdTAD-20354 317 1086 GGGccAGuuuGccAAGGAGdTdT 1598 CUCCUUGGcAAACUGGCCCdTdTAD-20355 318 1087 GGccAGuuuGccAAGGAGGdTdT 1599 CCUCCUUGGcAAACUGGCCdTdTAD-20356 319 1088 GccAGuuuGccAAGGAGGudTdT 1600 ACCUCCUUGGcAAACUGGCdTdTAD-20357 320 1089 ccAGuuuGccAAGGAGGuGdTdT 1601 cACCUCCUUGGcAAACUGGdTdTAD-20358 321 1090 cAGuuuGccAAGGAGGuGcdTdT 1602 GcACCUCCUUGGcAAACUGdTdTAD-20359 322 1091 AGuuuGccAAGGAGGuGcudTdT 1603 AGcACCUCCUUGGcAAACUdTdTAD-20360 323 1092 GuuuGccAAGGAGGuGcuGdTdT 1604 cAGcACCUCCUUGGcAAACdTdTAD-20361 324 1093 uuuGccAAGGAGGuGcuGcdTdT 1605 GcAGcACCUCCUUGGcAAAdTdTAD-20362 325 1094 uuGccAAGGAGGuGcuGccdTdT 1606 GGcAGcACCUCCUUGGcAAdTdTAD-20363 326 1095 uGccAAGGAGGuGcuGcccdTdT 1607 GGGcAGcACCUCCUUGGcAdTdTAD-20364 327 1096 GccAAGGAGGuGcuGcccAdTdT 1608 UGGGcAGcACCUCCUUGGCdTdTAD-20365 328 1097 ccAAGGAGGuGcuGcccAAdTdT 1609 UUGGGcAGcACCUCCUUGGdTdTAD-20366 329 1098 cAAGGAGGuGcuGcccAAGdTdT 1610 CUUGGGcAGcACCUCCUUGdTdTAD-20367 330 1099 AAGGAGGuGcuGcccAAGudTdT 1611 ACUUGGGcAGcACCUCCUUdTdTAD-20368 331 1100 AGGAGGuGcuGcccAAGuAdTdT 1612 uACUUGGGcAGcACCUCCUdTdTAD-20369 351 1101 uucAAGcAcAAcAAcAuGGdTdT 1613 CcAUGUUGUUGUGCUUGAAdTdTAD-20370 352 1102 ucAAGcAcAAcAAcAuGGcdTdT 1614 GCcAUGUUGUUGUGCUUGAdTdTAD-20371 353 1103 cAAGcAcAAcAAcAuGGccdTdT 1615 GGCcAUGUUGUUGUGCUUGdTdTAD-20372 354 1104 AAGcAcAAcAAcAuGGccAdTdT 1616 UGGCcAUGUUGUUGUGCUUdTdTAD-20373 355 1105 AGcAcAAcAAcAuGGccAGdTdT 1617 CUGGCcAUGUUGUUGUGCUdTdTAD-20374 356 1106 GcAcAAcAAcAuGGccAGcdTdT 1618 GCUGGCcAUGUUGUUGUGCdTdTAD-20375 357 1107 cAcAAcAAcAuGGccAGcudTdT 1619 AGCUGGCcAUGUUGUUGUGdTdTAD-20376 358 1108 AcAAcAAcAuGGccAGcuudTdT 1620 AAGCUGGCcAUGUUGUUGUdTdTAD-20378 360 1109 AAcAAcAuGGccAGcuucGdTdT 1621 CGAAGCUGGCcAUGUUGUUdTdTAD-20379 361 1110 AcAAcAuGGccAGcuucGudTdT 1622 ACGAAGCUGGCcAUGUUGUdTdTAD-20380 362 1111 cAAcAuGGccAGcuucGuGdTdT 1623 cACGAAGCUGGCcAUGUUGdTdTAD-20381 363 1112 AAcAuGGccAGcuucGuGcdTdT 1624 GcACGAAGCUGGCcAUGUUdTdTAD-20382 364 1113 AcAuGGccAGcuucGuGcGdTdT 1625 CGcACGAAGCUGGCcAUGUdTdTAD-20383 365 1114 cAuGGccAGcuucGuGcGGdTdT 1626 CCGcACGAAGCUGGCcAUGdTdTAD-20384 366 1115 AuGGccAGcuucGuGcGGcdTdT 1627 GCCGcACGAAGCUGGCcAUdTdTAD-20385 367 1116 uGGccAGcuucGuGcGGcAdTdT 1628 UGCCGcACGAAGCUGGCcAdTdTAD-20386 436 1117 uGGucAAGccAGAGAGAGAdTdT 1629 UCUCUCUCUGGCUUGACcAdTdTAD-20387 489 1139 GGccAGGAGcAGcuccuuGdTdT 1651 cAAGGAGCUGCUCCUGGCCdTdTAD-20388 490 1140 GccAGGAGcAGcuccuuGAdTdT 1652 UcAAGGAGCUGCUCCUGGCdTdTAD-20389 491 1141 ccAGGAGcAGcuccuuGAGdTdT 1653 CUcAAGGAGCUGCUCCUGGdTdTAD-20390 492 1142 cAGGAGcAGcuccuuGAGAdTdT 1654 UCUcAAGGAGCUGCUCCUGdTdTAD-20391 493 1143 AGGAGcAGcuccuuGAGAAdTdT 1655 UUCUcAAGGAGCUGCUCCUdTdTAD-20392 494 1144 GGAGcAGcuccuuGAGAAcdTdT 1656 GUUCUcAAGGAGCUGCUCCdTdTAD-20393 495 1145 GAGcAGcuccuuGAGAAcAdTdT 1657 UGUUCUcAAGGAGCUGCUCdTdTAD-20394 496 1146 AGcAGcuccuuGAGAAcAudTdT 1658 AUGUUCUcAAGGAGCUGCUdTdTAD-20395 497 1147 GcAGcuccuuGAGAAcAucdTdT 1659 GAUGUUCUcAAGGAGCUGCdTdTAD-20396 498 1148 cAGcuccuuGAGAAcAucAdTdT 1660 UGAUGUUCUcAAGGAGCUGdTdTAD-20397 499 1149 AGcuccuuGAGAAcAucAAdTdT 1661 UUGAUGUUCUcAAGGAGCUdTdTAD-20398 500 1150 GcuccuuGAGAAcAucAAGdTdT 1662 CUUGAUGUUCUcAAGGAGCdTdTAD-20399 501 1151 cuccuuGAGAAcAucAAGAdTdT 1663 UCUUGAUGUUCUcAAGGAGdTdTAD-20400 502 1152 uccuuGAGAAcAucAAGAGdTdT 1664 CUCUUGAUGUUCUcAAGGAdTdTAD-20401 503 1153 ccuuGAGAAcAucAAGAGGdTdT 1665 CCUCUUGAUGUUCUcAAGGdTdTAD-20402 504 1154 cuuGAGAAcAucAAGAGGAdTdT 1666 UCCUCUUGAUGUUCUcAAGdTdTAD-20403 505 1155 uuGAGAAcAucAAGAGGAAdTdT 1667 UUCCUCUUGAUGUUCUcAAdTdTAD-20404 506 1156 uGAGAAcAucAAGAGGAAAdTdT 1668 UUUCCUCUUGAUGUUCUcAdTdTAD-20405 509 1159 GAAcAucAAGAGGAAAGuGdTdT 1671 cACUUUCCUCUUGAUGUUCdTdTAD-20406 510 1160 AAcAucAAGAGGAAAGuGAdTdT 1672 UcACUUUCCUCUUGAUGUUdTdTAD-20407 511 1161 AcAucAAGAGGAAAGuGAcdTdT 1673 GUcACUUUCCUCUUGAUGUdTdTAD-20408 512 1162 cAucAAGAGGAAAGuGAccdTdT 1674 GGUcACUUUCCUCUUGAUGdTdTAD-20409 513 1163 AucAAGAGGAAAGuGAccAdTdT 1675 UGGUcACUUUCCUCUUGAUdTdTAD-20410 514 1164 ucAAGAGGAAAGuGAccAGdTdT 1676 CUGGUcACUUUCCUCUUGAdTdTAD-20411 515 1165 cAAGAGGAAAGuGAccAGudTdT 1677 ACUGGUcACUUUCCUCUUGdTdTAD-20412 516 1166 AAGAGGAAAGuGAccAGuGdTdT 1678 cACUGGUcACUUUCCUCUUdTdTAD-20413 517 2046 AGAGGAAAGuGAccAGuGudTdT 2047 AcACUGGUcACUUUCCUCUdTdTAD-20414 518 1167 GAGGAAAGuGAccAGuGuGdTdT 1679 cAcACUGGUcACUUUCCUCdTdTAD-20415 519 1168 AGGAAAGuGAccAGuGuGudTdT 1680 AcAcACUGGUcACUUUCCUdTdTAD-20416 520 1169 GGAAAGuGAccAGuGuGucdTdT 1681 GAcAcACUGGUcACUUUCCdTdTAD-20417 521 1170 GAAAGuGAccAGuGuGuccdTdT 1682 GGAcAcACUGGUcACUUUCdTdTAD-20418 522 1171 AAAGuGAccAGuGuGuccAdTdT 1683 UGGAcAcACUGGUcACUUUdTdTAD-20419 523 1172 AAGuGAccAGuGuGuccAcdTdT 1684 GUGGAcAcACUGGUcACUUdTdTAD-20420 524 1173 AGuGAccAGuGuGuccAccdTdT 1685 GGUGGAcAcACUGGUcACUdTdTAD-20421 525 1174 GuGAccAGuGuGuccAcccdTdT 1686 GGGUGGAcAcACUGGUcACdTdTAD-20422 526 1175 uGAccAGuGuGuccAcccudTdT 1687 AGGGUGGAcAcACUGGUcAdTdTAD-20423 527 1176 GAccAGuGuGuccAcccuGdTdT 1688 cAGGGUGGAcAcACUGGUCdTdTAD-20424 528 1177 AccAGuGuGuccAcccuGAdTdT 1689 UcAGGGUGGAcAcACUGGUdTdTAD-20425 529 1178 ccAGuGuGuccAcccuGAAdTdT 1690 UUcAGGGUGGAcAcACUGGdTdTAD-20426 530 1179 cAGuGuGuccAcccuGAAGdTdT 1691 CUUcAGGGUGGAcAcACUGdTdTAD-20427 531 1180 AGuGuGuccAcccuGAAGAdTdT 1692 UCUUcAGGGUGGAcAcACUdTdTAD-20428 532 1181 GuGuGuccAcccuGAAGAGdTdT 1693 CUCUUcAGGGUGGAcAcACdTdTAD-20429 533 1182 uGuGuccAcccuGAAGAGudTdT 1694 ACUCUUcAGGGUGGAcAcAdTdTAD-20430 534 1183 GuGuccAcccuGAAGAGuGdTdT 1695 cACUCUUcAGGGUGGAcACdTdTAD-20431 535 1184 uGuccAcccuGAAGAGuGAdTdT 1696 UcACUCUUcAGGGUGGAcAdTdTAD-20432 536 1185 GuccAcccuGAAGAGuGAAdTdT 1697 UUcACUCUUcAGGGUGGACdTdTAD-20433 537 1186 uccAcccuGAAGAGuGAAGdTdT 1698 CUUcACUCUUcAGGGUGGAdTdTAD-20434 538 1187 ccAcccuGAAGAGuGAAGAdTdT 1699 UCUUcACUCUUcAGGGUGGdTdTAD-20435 539 1188 cAcccuGAAGAGuGAAGAcdTdT 1700 GUCUUcACUCUUcAGGGUGdTdTAD-20436 540 1189 AcccuGAAGAGuGAAGAcAdTdT 1701 UGUCUUcACUCUUcAGGGUdTdTAD-20437 541 1190 cccuGAAGAGuGAAGAcAudTdT 1702 AUGUCUUcACUCUUcAGGGdTdTAD-20438 542 1191 ccuGAAGAGuGAAGAcAuAdTdT 1703 uAUGUCUUcACUCUUcAGGdTdTAD-20439 543 1192 cuGAAGAGuGAAGAcAuAAdTdT 1704 UuAUGUCUUcACUCUUcAGdTdTAD-20487 544 1193 uGAAGAGuGAAGAcAuAAAdTdT 1705 UUuAUGUCUUcACUCUUcAdTdTAD-20488 545 1194 GAAGAGuGAAGAcAuAAAGdTdT 1706 CUUuAUGUCUUcACUCUUCdTdTAD-20489 546 1195 AAGAGuGAAGAcAuAAAGAdTdT 1707 UCUUuAUGUCUUcACUCUUdTdTAD-20490 547 1196 AGAGuGAAGAcAuAAAGAudTdT 1708 AUCUUuAUGUCUUcACUCUdTdTAD-20491 548 1197 GAGuGAAGAcAuAAAGAucdTdT 1709 GAUCUUuAUGUCUUcACUCdTdTAD-20492 549 1198 AGuGAAGAcAuAAAGAuccdTdT 1710 GGAUCUUuAUGUCUUcACUdTdTAD-20493 550 1199 GuGAAGAcAuAAAGAuccGdTdT 1711 CGGAUCUUuAUGUCUUcACdTdTAD-20494 579 1200 GucAccAAGcuGcuGAcGGdTdT 1712 CCGUcAGcAGCUUGGUGACdTdTAD-20495 580 1201 ucAccAAGcuGcuGAcGGAdTdT 1713 UCCGUcAGcAGCUUGGUGAdTdTAD-20496 581 1202 cAccAAGcuGcuGAcGGAcdTdT 1714 GUCCGUcAGcAGCUUGGUGdTdTAD-20497 582 1203 AccAAGcuGcuGAcGGAcGdTdT 1715 CGUCCGUcAGcAGCUUGGUdTdTAD-20498 583 1204 ccAAGcuGcuGAcGGAcGudTdT 1716 ACGUCCGUcAGcAGCUUGGdTdTAD-20499 584 1205 cAAGcuGcuGAcGGAcGuGdTdT 1717 cACGUCCGUcAGcAGCUUGdTdTAD-20500 585 1206 AAGcuGcuGAcGGAcGuGcdTdT 1718 GcACGUCCGUcAGcAGCUUdTdTAD-20501 586 1207 AGcuGcuGAcGGAcGuGcAdTdT 1719 UGcACGUCCGUcAGcAGCUdTdTAD-20502 587 1208 GcuGcuGAcGGAcGuGcAGdTdT 1720 CUGcACGUCCGUcAGcAGCdTdTAD-20503 588 1209 cuGcuGAcGGAcGuGcAGcdTdT 1721 GCUGcACGUCCGUcAGcAGdTdTAD-20504 589 1210 uGcuGAcGGAcGuGcAGcudTdT 1722 AGCUGcACGUCCGUcAGcAdTdTAD-20505 590 1211 GcuGAcGGAcGuGcAGcuGdTdT 1723 cAGCUGcACGUCCGUcAGCdTdTAD-20506 591 1212 cuGAcGGAcGuGcAGcuGAdTdT 1724 UcAGCUGcACGUCCGUcAGdTdTAD-20507 592 1213 uGAcGGAcGuGcAGcuGAudTdT 1725 AUcAGCUGcACGUCCGUcAdTdTAD-20508 593 1214 GAcGGAcGuGcAGcuGAuGdTdT 1726 cAUcAGCUGcACGUCCGUCdTdTAD-20509 594 1215 AcGGAcGuGcAGcuGAuGAdTdT 1727 UcAUcAGCUGcACGUCCGUdTdTAD-20510 595 1216 cGGAcGuGcAGcuGAuGAAdTdT 1728 UUcAUcAGCUGcACGUCCGdTdTAD-20511 596 1217 GGAcGuGcAGcuGAuGAAGdTdT 1729 CUUcAUcAGCUGcACGUCCdTdTAD-20512 597 1218 GAcGuGcAGcuGAuGAAGGdTdT 1730 CCUUcAUcAGCUGcACGUCdTdTAD-20513 598 1219 AcGuGcAGcuGAuGAAGGGdTdT 1731 CCCUUcAUcAGCUGcACGUdTdTAD-20514 660 1220 GAGAAuGAGGcucuGuGGcdTdT 1732 GCcAcAGAGCCUcAUUCUCdTdTAD-20515 661 1221 AGAAuGAGGcucuGuGGcGdTdT 1733 CGCcAcAGAGCCUcAUUCUdTdTAD-20516 662 1222 GAAuGAGGcucuGuGGcGGdTdT 1734 CCGCcAcAGAGCCUcAUUCdTdTAD-20517 663 1223 AAuGAGGcucuGuGGcGGGdTdT 1735 CCCGCcAcAGAGCCUcAUUdTdTAD-20518 664 1224 AuGAGGcucuGuGGcGGGAdTdT 1736 UCCCGCcAcAGAGCCUcAUdTdTAD-20519 665 1225 uGAGGcucuGuGGcGGGAGdTdT 1737 CUCCCGCcAcAGAGCCUcAdTdTAD-20520 666 1226 GAGGcucuGuGGcGGGAGGdTdT 1738 CCUCCCGCcAcAGAGCCUCdTdTAD-20521 667 1227 AGGcucuGuGGcGGGAGGudTdT 1739 ACCUCCCGCcAcAGAGCCUdTdTAD-20522 668 1228 GGcucuGuGGcGGGAGGuGdTdT 1740 cACCUCCCGCcAcAGAGCCdTdTAD-20523 669 1229 GcucuGuGGcGGGAGGuGGdTdT 1741 CcACCUCCCGCcAcAGAGCdTdTAD-20524 670 1230 cucuGuGGcGGGAGGuGGcdTdT 1742 GCcACCUCCCGCcAcAGAGdTdTAD-20525 671 1231 ucuGuGGcGGGAGGuGGccdTdT 1743 GGCcACCUCCCGCcAcAGAdTdTAD-20526 672 1232 cuGuGGcGGGAGGuGGccAdTdT 1744 UGGCcACCUCCCGCcAcAGdTdTAD-20527 673 1233 uGuGGcGGGAGGuGGccAGdTdT 1745 CUGGCcACCUCCCGCcAcAdTdTAD-20528 674 1234 GuGGcGGGAGGuGGccAGcdTdT 1746 GCUGGCcACCUCCCGCcACdTdTAD-20529 675 1235 uGGcGGGAGGuGGccAGccdTdT 1747 GGCUGGCcACCUCCCGCcAdTdTAD-20530 676 1236 GGcGGGAGGuGGccAGccudTdT 1748 AGGCUGGCcACCUCCCGCCdTdTAD-20531 677 1237 GcGGGAGGuGGccAGccuudTdT 1749 AAGGCUGGCcACCUCCCGCdTdTAD-20532 678 1238 cGGGAGGuGGccAGccuucdTdT 1750 GAAGGCUGGCcACCUCCCGdTdTAD-20533 679 1239 GGGAGGuGGccAGccuucGdTdT 1751 CGAAGGCUGGCcACCUCCCdTdTAD-20534 680 1240 GGAGGuGGccAGccuucGGdTdT 1752 CCGAAGGCUGGCcACCUCCdTdTAD-20535 681 1241 GAGGuGGccAGccuucGGcdTdT 1753 GCCGAAGGCUGGCcACCUCdTdTAD-20536 682 1242 AGGuGGccAGccuucGGcAdTdT 1754 UGCCGAAGGCUGGCcACCUdTdTAD-20537 683 1243 GGuGGccAGccuucGGcAGdTdT 1755 CUGCCGAAGGCUGGCcACCdTdTAD-20538 684 1244 GuGGccAGccuucGGcAGAdTdT 1756 UCUGCCGAAGGCUGGCcACdTdTAD-20539 685 1245 uGGccAGccuucGGcAGAAdTdT 1757 UUCUGCCGAAGGCUGGCcAdTdTAD-20540 690 1250 AGccuucGGcAGAAGcAuGdTdT 1762 cAUGCUUCUGCCGAAGGCUdTdTAD-20541 691 1251 GccuucGGcAGAAGcAuGcdTdT 1763 GcAUGCUUCUGCCGAAGGCdTdTAD-20542 692 1252 ccuucGGcAGAAGcAuGccdTdT 1764 GGcAUGCUUCUGCCGAAGGdTdTAD-20543 693 1253 cuucGGcAGAAGcAuGcccdTdT 1765 GGGcAUGCUUCUGCCGAAGdTdTAD-20544 694 1254 uucGGcAGAAGcAuGcccAdTdT 1766 UGGGcAUGCUUCUGCCGAAdTdTAD-20545 695 1255 ucGGcAGAAGcAuGcccAGdTdT 1767 CUGGGcAUGCUUCUGCCGAdTdTAD-20546 696 1256 cGGcAGAAGcAuGcccAGcdTdT 1768 GCUGGGcAUGCUUCUGCCGdTdTAD-20547 697 1257 GGcAGAAGcAuGcccAGcAdTdT 1769 UGCUGGGcAUGCUUCUGCCdTdTAD-20548 698 1258 GcAGAAGcAuGcccAGcAAdTdT 1770 UUGCUGGGcAUGCUUCUGCdTdTAD-20549 699 1259 cAGAAGcAuGcccAGcAAcdTdT 1771 GUUGCUGGGcAUGCUUCUGdTdTAD-20550 700 1260 AGAAGcAuGcccAGcAAcAdTdT 1772 UGUUGCUGGGcAUGCUUCUdTdTAD-20551 701 1261 GAAGcAuGcccAGcAAcAGdTdT 1773 CUGUUGCUGGGcAUGCUUCdTdTAD-20552 702 1262 AAGcAuGcccAGcAAcAGAdTdT 1774 UCUGUUGCUGGGcAUGCUUdTdTAD-20553 703 1263 AGcAuGcccAGcAAcAGAAdTdT 1775 UUCUGUUGCUGGGcAUGCUdTdTAD-20554 704 1264 GcAuGcccAGcAAcAGAAAdTdT 1776 UUUCUGUUGCUGGGcAUGCdTdTAD-20555 705 1265 cAuGcccAGcAAcAGAAAGdTdT 1777 CUUUCUGUUGCUGGGcAUGdTdTAD-20556 706 1266 AuGcccAGcAAcAGAAAGudTdT 1778 ACUUUCUGUUGCUGGGcAUdTdTAD-20557 707 1267 uGcccAGcAAcAGAAAGucdTdT 1779 GACUUUCUGUUGCUGGGcAdTdTAD-20558 731 1291 cAAGcucAuucAGuuccuGdTdT 1803 cAGGAACUGAAUGAGCUUGdTdTAD-20559 732 1292 AAGcucAuucAGuuccuGAdTdT 1804 UcAGGAACUGAAUGAGCUUdTdTAD-20560 733 1293 AGcucAuucAGuuccuGAudTdT 1805 AUcAGGAACUGAAUGAGCUdTdTAD-20561 734 1294 GcucAuucAGuuccuGAucdTdT 1806 GAUcAGGAACUGAAUGAGCdTdTAD-20562 735 1295 cucAuucAGuuccuGAucudTdT 1807 AGAUcAGGAACUGAAUGAGdTdTAD-20563 736 1296 ucAuucAGuuccuGAucucdTdT 1808 GAGAUcAGGAACUGAAUGAdTdTAD-20564 737 1297 cAuucAGuuccuGAucucAdTdT 1809 UGAGAUcAGGAACUGAAUGdTdTAD-20565 738 1298 AuucAGuuccuGAucucAcdTdT 1810 GUGAGAUcAGGAACUGAAUdTdTAD-20566 739 1299 uucAGuuccuGAucucAcudTdT 1811 AGUGAGAUcAGGAACUGAAdTdTAD-20567 740 1300 ucAGuuccuGAucucAcuGdTdT 1812 cAGUGAGAUcAGGAACUGAdTdTAD-20568 741 1301 cAGuuccuGAucucAcuGGdTdT 1813 CcAGUGAGAUcAGGAACUGdTdTAD-20569 742 1302 AGuuccuGAucucAcuGGudTdT 1814 ACcAGUGAGAUcAGGAACUdTdTAD-20571 744 1303 uuccuGAucucAcuGGuGcdTdT 1815 GcACcAGUGAGAUcAGGAAdTdTAD-20572 745 1304 uccuGAucucAcuGGuGcAdTdT 1816 UGcACcAGUGAGAUcAGGAdTdTAD-20573 746 1305 ccuGAucucAcuGGuGcAGdTdT 1817 CUGcACcAGUGAGAUcAGGdTdTAD-20574 747 2048 cuGAucucAcuGGuGcAGudTdT 2049 ACUGcACcAGUGAGAUcAGdTdTAD-20575 748 1306 uGAucucAcuGGuGcAGucdTdT 1818 GACUGcACcAGUGAGAUcAdTdTAD-20576 749 1307 GAucucAcuGGuGcAGucAdTdT 1819 UGACUGcACcAGUGAGAUCdTdTAD-20577 750 1308 AucucAcuGGuGcAGucAAdTdT 1820 UUGACUGcACcAGUGAGAUdTdTAD-20578 751 1309 ucucAcuGGuGcAGucAAAdTdT 1821 UUUGACUGcACcAGUGAGAdTdTAD-20579 752 1310 cucAcuGGuGcAGucAAAcdTdT 1822 GUUUGACUGcACcAGUGAGdTdTAD-20581 754 1311 cAcuGGuGcAGucAAAccGdTdT 1823 CGGUUUGACUGcACcAGUGdTdTAD-20582 755 1312 AcuGGuGcAGucAAAccGGdTdT 1824 CCGGUUUGACUGcACcAGUdTdTAD-20625 756 1313 cuGGuGcAGucAAAccGGAdTdT 1825 UCCGGUUUGACUGcACcAGdTdTAD-20626 757 1314 uGGuGcAGucAAAccGGAudTdT 1826 AUCCGGUUUGACUGcACcAdTdTAD-20627 758 1315 GGuGcAGucAAAccGGAucdTdT 1827 GAUCCGGUUUGACUGcACCdTdTAD-20628 759 1316 GuGcAGucAAAccGGAuccdTdT 1828 GGAUCCGGUUUGACUGcACdTdTAD-20629 760 1317 uGcAGucAAAccGGAuccudTdT 1829 AGGAUCCGGUUUGACUGcAdTdTAD-20630 761 1318 GcAGucAAAccGGAuccuGdTdT 1830 cAGGAUCCGGUUUGACUGCdTdTAD-20631 762 1319 cAGucAAAccGGAuccuGGdTdT 1831 CcAGGAUCCGGUUUGACUGdTdTAD-20632 763 1320 AGucAAAccGGAuccuGGGdTdT 1832 CCcAGGAUCCGGUUUGACUdTdTAD-20633 781 1321 GGGuGAAGAGAAAGAucccdTdT 1833 GGGAUCUUUCUCUUcACCCdTdTAD-20634 799 1322 cccuGAuGcuGAAcGAcAGdTdT 1834 CUGUCGUUcAGcAUcAGGGdTdTAD-20635 800 1323 ccuGAuGcuGAAcGAcAGudTdT 1835 ACUGUCGUUcAGcAUcAGGdTdTAD-20636 801 1324 cuGAuGcuGAAcGAcAGuGdTdT 1836 cACUGUCGUUcAGcAUcAGdTdTAD-20637 802 1325 uGAuGcuGAAcGAcAGuGGdTdT 1837 CcACUGUCGUUcAGcAUcAdTdTAD-20638 803 1326 GAuGcuGAAcGAcAGuGGcdTdT 1838 GCcACUGUCGUUcAGcAUCdTdTAD-20639 804 1327 AuGcuGAAcGAcAGuGGcudTdT 1839 AGCcACUGUCGUUcAGcAUdTdTAD-20640 805 1328 uGcuGAAcGAcAGuGGcucdTdT 1840 GAGCcACUGUCGUUcAGcAdTdTAD-20641 806 1329 GcuGAAcGAcAGuGGcucAdTdT 1841 UGAGCcACUGUCGUUcAGCdTdTAD-20642 807 1330 cuGAAcGAcAGuGGcucAGdTdT 1842 CUGAGCcACUGUCGUUcAGdTdTAD-20643 808 1331 uGAAcGAcAGuGGcucAGcdTdT 1843 GCUGAGCcACUGUCGUUcAdTdTAD-20644 809 1332 GAAcGAcAGuGGcucAGcAdTdT 1844 UGCUGAGCcACUGUCGUUCdTdTAD-20645 810 1333 AAcGAcAGuGGcucAGcAcdTdT 1845 GUGCUGAGCcACUGUCGUUdTdTAD-20646 811 1334 AcGAcAGuGGcucAGcAcAdTdT 1846 UGUGCUGAGCcACUGUCGUdTdTAD-20647 812 1335 cGAcAGuGGcucAGcAcAudTdT 1847 AUGUGCUGAGCcACUGUCGdTdTAD-20648 813 1336 GAcAGuGGcucAGcAcAuudTdT 1848 AAUGUGCUGAGCcACUGUCdTdTAD-20649 814 1337 AcAGuGGcucAGcAcAuucdTdT 1849 GAAUGUGCUGAGCcACUGUdTdTAD-20650 815 1338 cAGuGGcucAGcAcAuuccdTdT 1850 GGAAUGUGCUGAGCcACUGdTdTAD-20651 816 1339 AGuGGcucAGcAcAuuccAdTdT 1851 UGGAAUGUGCUGAGCcACUdTdTAD-20652 817 1340 GuGGcucAGcAcAuuccAudTdT 1852 AUGGAAUGUGCUGAGCcACdTdTAD-20653 818 1341 uGGcucAGcAcAuuccAuGdTdT 1853 cAUGGAAUGUGCUGAGCcAdTdTAD-20654 819 1342 GGcucAGcAcAuuccAuGcdTdT 1854 GcAUGGAAUGUGCUGAGCCdTdTAD-20655 820 1343 GcucAGcAcAuuccAuGccdTdT 1855 GGcAUGGAAUGUGCUGAGCdTdTAD-20656 821 1344 cucAGcAcAuuccAuGcccdTdT 1856 GGGcAUGGAAUGUGCUGAGdTdTAD-20657 822 1345 ucAGcAcAuuccAuGcccAdTdT 1857 UGGGcAUGGAAUGUGCUGAdTdTAD-20658 823 1346 cAGcAcAuuccAuGcccAAdTdT 1858 UUGGGcAUGGAAUGUGCUGdTdTAD-20659 824 1347 AGcAcAuuccAuGcccAAGdTdT 1859 CUUGGGcAUGGAAUGUGCUdTdTAD-20660 825 1348 GcAcAuuccAuGcccAAGudTdT 1860 ACUUGGGcAUGGAAUGUGCdTdTAD-20661 826 1349 cAcAuuccAuGcccAAGuAdTdT 1861 uACUUGGGcAUGGAAUGUGdTdTAD-20284 827 1350 AcAuuccAuGcccAAGuAudTdT 1862 AuACUUGGGcAUGGAAUGUdTdTAD-20662 847 1351 GccGGcAGuucucccuGGAdTdT 1863 UCcAGGGAGAACUGCCGGCdTdTAD-20868 848 1352 ccGGcAGuucucccuGGAGdTdT 1864 CUCcAGGGAGAACUGCCGGdTdTAD-20663 849 1353 cGGcAGuucucccuGGAGcdTdT 1865 GCUCcAGGGAGAACUGCCGdTdTAD-20664 850 1354 GGcAGuucucccuGGAGcAdTdT 1866 UGCUCcAGGGAGAACUGCCdTdTAD-20665 851 1355 GcAGuucucccuGGAGcAcdTdT 1867 GUGCUCcAGGGAGAACUGCdTdTAD-20666 852 1356 cAGuucucccuGGAGcAcGdTdT 1868 CGUGCUCcAGGGAGAACUGdTdTAD-20667 853 1357 AGuucucccuGGAGcAcGudTdT 1869 ACGUGCUCcAGGGAGAACUdTdTAD-20668 854 1358 GuucucccuGGAGcAcGucdTdT 1870 GACGUGCUCcAGGGAGAACdTdTAD-20669 855 1359 uucucccuGGAGcAcGuccdTdT 1871 GGACGUGCUCcAGGGAGAAdTdTAD-20670 856 1360 ucucccuGGAGcAcGuccAdTdT 1872 UGGACGUGCUCcAGGGAGAdTdTAD-20671 857 1361 cucccuGGAGcAcGuccAcdTdT 1873 GUGGACGUGCUCcAGGGAGdTdTAD-20672 858 1362 ucccuGGAGcAcGuccAcGdTdT 1874 CGUGGACGUGCUCcAGGGAdTdTAD-20673 859 1363 cccuGGAGcAcGuccAcGGdTdT 1875 CCGUGGACGUGCUCcAGGGdTdTAD-20674 860 1364 ccuGGAGcAcGuccAcGGcdTdT 1876 GCCGUGGACGUGCUCcAGGdTdTAD-20675 861 1365 cuGGAGcAcGuccAcGGcudTdT 1877 AGCCGUGGACGUGCUCcAGdTdTAD-20676 862 1366 uGGAGcAcGuccAcGGcucdTdT 1878 GAGCCGUGGACGUGCUCcAdTdTAD-20677 915 1373 AGcuccAGccucuAcGcccdTdT 1885 GGGCGuAGAGGCUGGAGCUdTdTAD-20678 965 1385 cuccGAcAucAccGAGcuGdTdT 1897 cAGCUCGGUGAUGUCGGAGdTdTAD-20679 966 1386 uccGAcAucAccGAGcuGGdTdT 1898 CcAGCUCGGUGAUGUCGGAdTdTAD-20680 967 1387 ccGAcAucAccGAGcuGGcdTdT 1899 GCcAGCUCGGUGAUGUCGGdTdTAD-20681 968 1388 cGAcAucAccGAGcuGGcudTdT 1900 AGCcAGCUCGGUGAUGUCGdTdTAD-20682 969 1389 GAcAucAccGAGcuGGcucdTdT 1901 GAGCcAGCUCGGUGAUGUCdTdTAD-20683 970 1390 AcAucAccGAGcuGGcuccdTdT 1902 GGAGCcAGCUCGGUGAUGUdTdTAD-20684 971 1391 cAucAccGAGcuGGcuccudTdT 1903 AGGAGCcAGCUCGGUGAUGdTdTAD-20685 972 1392 AucAccGAGcuGGcuccuGdTdT 1904 cAGGAGCcAGCUCGGUGAUdTdTAD-20686 973 1393 ucAccGAGcuGGcuccuGcdTdT 1905 GcAGGAGCcAGCUCGGUGAdTdTAD-20687 974 1394 cAccGAGcuGGcuccuGccdTdT 1906 GGcAGGAGCcAGCUCGGUGdTdTAD-20688 975 1395 AccGAGcuGGcuccuGccAdTdT 1907 UGGcAGGAGCcAGCUCGGUdTdTAD-20689 976 1396 ccGAGcuGGcuccuGccAGdTdT 1908 CUGGcAGGAGCcAGCUCGGdTdTAD-20690 977 1397 cGAGcuGGcuccuGccAGcdTdT 1909 GCUGGcAGGAGCcAGCUCGdTdTAD-20691 978 1398 GAGcuGGcuccuGccAGccdTdT 1910 GGCUGGcAGGAGCcAGCUCdTdTAD-20692 979 1399 AGcuGGcuccuGccAGcccdTdT 1911 GGGCUGGcAGGAGCcAGCUdTdTAD-20693 1011 1400 GGcGGGAGcAuAGAcGAGAdTdT 1912 UCUCGUCuAUGCUCCCGCCdTdTAD-20694 1012 1401 GcGGGAGcAuAGAcGAGAGdTdT 1913 CUCUCGUCuAUGCUCCCGCdTdTAD-20695 1013 1402 cGGGAGcAuAGAcGAGAGGdTdT 1914 CCUCUCGUCuAUGCUCCCGdTdTAD-20696 1014 1403 GGGAGcAuAGAcGAGAGGcdTdT 1915 GCCUCUCGUCuAUGCUCCCdTdTAD-20697 1015 1404 GGAGcAuAGAcGAGAGGccdTdT 1916 GGCCUCUCGUCuAUGCUCCdTdTAD-20698 1016 1405 GAGcAuAGAcGAGAGGcccdTdT 1917 GGGCCUCUCGUCuAUGCUCdTdTAD-20699 1048 1406 cccuGGuGcGuGucAAGGAdTdT 1918 UCCUUGAcACGcACcAGGGdTdTAD-20700 1049 1407 ccuGGuGcGuGucAAGGAGdTdT 1919 CUCCUUGAcACGcACcAGGdTdTAD-20701 1050 1408 cuGGuGcGuGucAAGGAGGdTdT 1920 CCUCCUUGAcACGcACcAGdTdTAD-20702 1051 1409 uGGuGcGuGucAAGGAGGAdTdT 1921 UCCUCCUUGAcACGcACcAdTdTAD-20869 1052 1410 GGuGcGuGucAAGGAGGAGdTdT 1922 CUCCUCCUUGAcACGcACCdTdTAD-20703 1053 1411 GuGcGuGucAAGGAGGAGcdTdT 1923 GCUCCUCCUUGAcACGcACdTdTAD-20704 1054 1412 uGcGuGucAAGGAGGAGccdTdT 1924 GGCUCCUCCUUGAcACGcAdTdTAD-20705 1055 1413 GcGuGucAAGGAGGAGcccdTdT 1925 GGGCUCCUCCUUGAcACGCdTdTAD-20706 1329 1416 cAcuuGGAuGcuAuGGAcudTdT 1928 AGUCcAuAGcAUCcAAGUGdTdTAD-20707 1330 1417 AcuuGGAuGcuAuGGAcucdTdT 1929 GAGUCcAuAGcAUCcAAGUdTdTAD-20709 1332 1418 uuGGAuGcuAuGGAcuccAdTdT 1930 UGGAGUCcAuAGcAUCcAAdTdTAD-20710 1333 1419 uGGAuGcuAuGGAcuccAAdTdT 1931 UUGGAGUCcAuAGcAUCcAdTdTAD-20711 1334 1420 GGAuGcuAuGGAcuccAAcdTdT 1932 GUUGGAGUCcAuAGcAUCCdTdTAD-20712 1335 1421 GAuGcuAuGGAcuccAAccdTdT 1933 GGUUGGAGUCcAuAGcAUCdTdTAD-20713 1336 1422 AuGcuAuGGAcuccAAccudTdT 1934 AGGUUGGAGUCcAuAGcAUdTdTAD-20714 1337 1423 uGcuAuGGAcuccAAccuGdTdT 1935 cAGGUUGGAGUCcAuAGcAdTdTAD-20715 1338 1424 GcuAuGGAcuccAAccuGGdTdT 1936 CcAGGUUGGAGUCcAuAGCdTdTAD-20716 1339 1425 cuAuGGAcuccAAccuGGAdTdT 1937 UCcAGGUUGGAGUCcAuAGdTdTAD-20717 1359 1426 AAccuGcAGAccAuGcuGAdTdT 1938 UcAGcAUGGUCUGcAGGUUdTdTAD-20718 1360 1427 AccuGcAGAccAuGcuGAGdTdT 1939 CUcAGcAUGGUCUGcAGGUdTdTAD-20719 1361 1428 ccuGcAGAccAuGcuGAGcdTdT 1940 GCUcAGcAUGGUCUGcAGGdTdTAD-20720 1362 1429 cuGcAGAccAuGcuGAGcAdTdT 1941 UGCUcAGcAUGGUCUGcAGdTdTAD-20721 1363 1430 uGcAGAccAuGcuGAGcAGdTdT 1942 CUGCUcAGcAUGGUCUGcAdTdTAD-20722 1364 1431 GcAGAccAuGcuGAGcAGcdTdT 1943 GCUGCUcAGcAUGGUCUGCdTdTAD-20723 1365 1432 cAGAccAuGcuGAGcAGccdTdT 1944 GGCUGCUcAGcAUGGUCUGdTdTAD-20724 1366 1433 AGAccAuGcuGAGcAGccAdTdT 1945 UGGCUGCUcAGcAUGGUCUdTdTAD-20725 1367 1434 GAccAuGcuGAGcAGccAcdTdT 1946 GUGGCUGCUcAGcAUGGUCdTdTAD-20726 1368 1435 AccAuGcuGAGcAGccAcGdTdT 1947 CGUGGCUGCUcAGcAUGGUdTdTAD-20727 1369 1436 ccAuGcuGAGcAGccAcGGdTdT 1948 CCGUGGCUGCUcAGcAUGGdTdTAD-20728 1370 1437 cAuGcuGAGcAGccAcGGcdTdT 1949 GCCGUGGCUGCUcAGcAUGdTdTAD-20729 1371 1438 AuGcuGAGcAGccAcGGcudTdT 1950 AGCCGUGGCUGCUcAGcAUdTdTAD-20730 1372 1439 uGcuGAGcAGccAcGGcuudTdT 1951 AAGCCGUGGCUGCUcAGcAdTdTAD-20731 1373 1440 GcuGAGcAGccAcGGcuucdTdT 1952 GAAGCCGUGGCUGCUcAGCdTdTAD-20732 1374 1441 cuGAGcAGccAcGGcuucAdTdT 1953 UGAAGCCGUGGCUGCUcAGdTdTAD-20733 1375 1442 uGAGcAGccAcGGcuucAGdTdT 1954 CUGAAGCCGUGGCUGCUcAdTdTAD-20734 1379 1446 cAGccAcGGcuucAGcGuGdTdT 1958 cACGCUGAAGCCGUGGCUGdTdTAD-20735 1380 1447 AGccAcGGcuucAGcGuGGdTdT 1959 CcACGCUGAAGCCGUGGCUdTdTAD-20736 1381 1448 GccAcGGcuucAGcGuGGAdTdT 1960 UCcACGCUGAAGCCGUGGCdTdTAD-20737 1382 1449 ccAcGGcuucAGcGuGGAcdTdT 1961 GUCcACGCUGAAGCCGUGGdTdTAD-20738 1383 1450 cAcGGcuucAGcGuGGAcAdTdT 1962 UGUCcACGCUGAAGCCGUGdTdTAD-20739 1384 1451 AcGGcuucAGcGuGGAcAcdTdT 1963 GUGUCcACGCUGAAGCCGUdTdTAD-20740 1385 1452 cGGcuucAGcGuGGAcAccdTdT 1964 GGUGUCcACGCUGAAGCCGdTdTAD-20741 1386 1453 GGcuucAGcGuGGAcAccAdTdT 1965 UGGUGUCcACGCUGAAGCCdTdTAD-20742 1387 1454 GcuucAGcGuGGAcAccAGdTdT 1966 CUGGUGUCcACGCUGAAGCdTdTAD-20743 1407 1455 GcccuGcuGGAccuGuucAdTdT 1967 UGAAcAGGUCcAGcAGGGCdTdTAD-20744 1408 1456 cccuGcuGGAccuGuucAGdTdT 1968 CUGAAcAGGUCcAGcAGGGdTdTAD-20745 1409 1457 ccuGcuGGAccuGuucAGcdTdT 1969 GCUGAAcAGGUCcAGcAGGdTdTAD-20746 1410 1458 cuGcuGGAccuGuucAGccdTdT 1970 GGCUGAAcAGGUCcAGcAGdTdTAD-20747 1411 1459 uGcuGGAccuGuucAGcccdTdT 1971 GGGCUGAAcAGGUCcAGcAdTdTAD-20748 1428 1460 cccucGGuGAccGuGcccGdTdT 1972 CGGGcACGGUcACCGAGGGdTdTAD-20749 1429 1461 ccucGGuGAccGuGcccGAdTdT 1973 UCGGGcACGGUcACCGAGGdTdTAD-20750 1430 1462 cucGGuGAccGuGcccGAcdTdT 1974 GUCGGGcACGGUcACCGAGdTdTAD-20751 1431 1463 ucGGuGAccGuGcccGAcAdTdT 1975 UGUCGGGcACGGUcACCGAdTdTAD-20752 1432 1464 cGGuGAccGuGcccGAcAudTdT 1976 AUGUCGGGcACGGUcACCGdTdTAD-20753 1433 1465 GGuGAccGuGcccGAcAuGdTdT 1977 cAUGUCGGGcACGGUcACCdTdTAD-20754 1434 1466 GuGAccGuGcccGAcAuGAdTdT 1978 UcAUGUCGGGcACGGUcACdTdTAD-20755 1435 1467 uGAccGuGcccGAcAuGAGdTdT 1979 CUcAUGUCGGGcACGGUcAdTdTAD-20756 1436 1468 GAccGuGcccGAcAuGAGcdTdT 1980 GCUcAUGUCGGGcACGGUCdTdTAD-20757 1437 1469 AccGuGcccGAcAuGAGccdTdT 1981 GGCUcAUGUCGGGcACGGUdTdTAD-20758 1438 1470 ccGuGcccGAcAuGAGccudTdT 1982 AGGCUcAUGUCGGGcACGGdTdTAD-20759 1439 1471 cGuGcccGAcAuGAGccuGdTdT 1983 cAGGCUcAUGUCGGGcACGdTdTAD-20760 1440 1472 GuGcccGAcAuGAGccuGcdTdT 1984 GcAGGCUcAUGUCGGGcACdTdTAD-20761 1441 1473 uGcccGAcAuGAGccuGccdTdT 1985 GGcAGGCUcAUGUCGGGcAdTdTAD-20762 1442 1474 GcccGAcAuGAGccuGccudTdT 1986 AGGcAGGCUcAUGUCGGGCdTdTAD-20763 1443 1475 cccGAcAuGAGccuGccuGdTdT 1987 cAGGcAGGCUcAUGUCGGGdTdTAD-20764 1444 1476 ccGAcAuGAGccuGccuGAdTdT 1988 UcAGGcAGGCUcAUGUCGGdTdTAD-20765 1445 1477 cGAcAuGAGccuGccuGAcdTdT 1989 GUcAGGcAGGCUcAUGUCGdTdTAD-20766 1446 1478 GAcAuGAGccuGccuGAccdTdT 1990 GGUcAGGcAGGCUcAUGUCdTdTAD-20767 1447 1479 AcAuGAGccuGccuGAccudTdT 1991 AGGUcAGGcAGGCUcAUGUdTdTAD-20768 1448 1480 cAuGAGccuGccuGAccuudTdT 1992 AAGGUcAGGcAGGCUcAUGdTdTAD-20769 1449 1481 AuGAGccuGccuGAccuuGdTdT 1993 cAAGGUcAGGcAGGCUcAUdTdTAD-20770 1450 1482 uGAGccuGccuGAccuuGAdTdT 1994 UcAAGGUcAGGcAGGCUcAdTdTAD-20771 1451 1483 GAGccuGccuGAccuuGAcdTdT 1995 GUcAAGGUcAGGcAGGCUCdTdTAD-20772 1452 1484 AGccuGccuGAccuuGAcAdTdT 1996 UGUcAAGGUcAGGcAGGCUdTdTAD-20773 1453 1485 GccuGccuGAccuuGAcAGdTdT 1997 CUGUcAAGGUcAGGcAGGCdTdTAD-20774 1454 1486 ccuGccuGAccuuGAcAGcdTdT 1998 GCUGUcAAGGUcAGGcAGGdTdTAD-20775 1455 1487 cuGccuGAccuuGAcAGcAdTdT 1999 UGCUGUcAAGGUcAGGcAGdTdTAD-20776 1456 1488 uGccuGAccuuGAcAGcAGdTdT 2000 CUGCUGUcAAGGUcAGGcAdTdTAD-20777 1457 1489 GccuGAccuuGAcAGcAGcdTdT 2001 GCUGCUGUcAAGGUcAGGCdTdTAD-20778 1458 1490 ccuGAccuuGAcAGcAGccdTdT 2002 GGCUGCUGUcAAGGUcAGGdTdTAD-20779 1459 1491 cuGAccuuGAcAGcAGccudTdT 2003 AGGCUGCUGUcAAGGUcAGdTdTAD-20780 1460 1492 uGAccuuGAcAGcAGccuGdTdT 2004 cAGGCUGCUGUcAAGGUcAdTdTAD-20781 1461 1493 GAccuuGAcAGcAGccuGGdTdT 2005 CcAGGCUGCUGUcAAGGUCdTdTAD-20782 1462 1494 AccuuGAcAGcAGccuGGcdTdT 2006 GCcAGGCUGCUGUcAAGGUdTdTAD-20783 1482 1495 AGuAuccAAGAGcuccuGudTdT 2007 AcAGGAGCUCUUGGAuACUdTdTAD-20784 1483 1496 GuAuccAAGAGcuccuGucdTdT 2008 GAcAGGAGCUCUUGGAuACdTdTAD-20785 1484 1497 uAuccAAGAGcuccuGucudTdT 2009 AGAcAGGAGCUCUUGGAuAdTdTAD-20786 1485 1498 AuccAAGAGcuccuGucucdTdT 2010 GAGAcAGGAGCUCUUGGAUdTdTAD-20787 1486 1499 uccAAGAGcuccuGucuccdTdT 2011 GGAGAcAGGAGCUCUUGGAdTdTAD-20788 1487 1500 ccAAGAGcuccuGucucccdTdT 2012 GGGAGAcAGGAGCUCUUGGdTdTAD-20789 1547 1515 GGAuucAGGGAAGcAGcuGdTdT 2027 cAGCUGCUUCCCUGAAUCCdTdTAD-20790 1548 1516 GAuucAGGGAAGcAGcuGGdTdT 2028 CcAGCUGCUUCCCUGAAUCdTdTAD-20791 1549 1517 AuucAGGGAAGcAGcuGGudTdT 2029 ACcAGCUGCUUCCCUGAAUdTdTAD-20792 1602 1518 cccGGcuccGuGGAcAccGdTdT 2030 CGGUGUCcACGGAGCCGGGdTdTAD-20793 1603 1519 ccGGcuccGuGGAcAccGGdTdT 2031 CCGGUGUCcACGGAGCCGGdTdTAD-20794 1604 1520 cGGcuccGuGGAcAccGGGdTdT 2032 CCCGGUGUCcACGGAGCCGdTdTAD-20795 1605 1521 GGcuccGuGGAcAccGGGAdTdT 2033 UCCCGGUGUCcACGGAGCCdTdTAD-20796 1606 1522 GcuccGuGGAcAccGGGAGdTdT 2034 CUCCCGGUGUCcACGGAGCdTdTAD-20870 1607 1523 cuccGuGGAcAccGGGAGcdTdT 2035 GCUCCCGGUGUCcACGGAGdTdTAD-20871 1608 1524 uccGuGGAcAccGGGAGcAdTdT 2036 UGCUCCCGGUGUCcACGGAdTdTAD-20872 1633 1525 uGccGGuGcuGuuuGAGcudTdT 2037 AGCUcAAAcAGcACCGGcAdTdTAD-20797 1634 1526 GccGGuGcuGuuuGAGcuGdTdT 2038 cAGCUcAAAcAGcACCGGCdTdTAD-20798 1635 1527 ccGGuGcuGuuuGAGcuGGdTdT 2039 CcAGCUcAAAcAGcACCGGdTdTAD-20799 1636 1528 cGGuGcuGuuuGAGcuGGGdTdT 2040 CCcAGCUcAAAcAGcACCGdTdTAD-20873 1698 1529 cccAccAucucccuGcuGAdTdT 2041 UcAGcAGGGAGAUGGUGGGdTdTAD-20800 1699 1530 ccAccAucucccuGcuGAcdTdT 3282 GUcAGcAGGGAGAUGGUGGdTdTAD-20801 1700 1531 cAccAucucccuGcuGAcAdTdT 3283 UGUcAGcAGGGAGAUGGUGdTdTAD-20278 2075 GcAGGuuGuucAuAGucAGdTdT 2086 CUGACuAUGAAcAACCUGCdTdTAD-20279 2076 cAGGuuGuucAuAGucAGAdTdT 2087 UCUGACuAUGAAcAACCUGdTdTAD-20280 2077 AGGuuGuucAuAGucAGAAdTdT 2088 UUCUGACuAUGAAcAACCUdTdTAD-20281 2078 GcccAAGuAcuucAAGcAcdTdT 2089 GUGCUUGAAGuACUUGGGCdTdTAD-20282 2079 cccAAGuAcuucAAGcAcAdTdT 2090 UGUGCUUGAAGuACUUGGGdTdTAD-20283 2080 ccAAGuAcuucAAGcAcAAdTdT 2091 UUGUGCUUGAAGuACUUGGdTdTAD-20377 2081 cAAcAAcAuGGccAGcuucdTdT 2092 GAAGCUGGCcAUGUUGUUGdTdTAD-20570 2082 GuuccuGAucucAcuGGuGdTdT 2093 cACcAGUGAGAUcAGGAACdTdTAD-20580 2083 ucAcuGGuGcAGucAAAccdTdT 2094 GGUUUGACUGcACcAGUGAdTdTAD-20597 2084 UUGGUcAGGAAGGCCGGGAdTdT 2095 ucccGGccuuccuGAccAAdTdTAD-20598 2085 cccGGccuuccuGAccAAGdTdT 2096 CUUGGUcAGGAAGGCCGGGdTdT

Abbreviations in the nucleotide sequences herein are as depicted inTable 2A.

TABLE 2A ABBREVIATIONS Abbreviation Nucleotide(s) Aadenosine-5′-phosphate C cytidine-5′-phosphate G guanosine-5′-phosphatedT 2′-deoxy-thymidine-5′-phosphate U uridine-5′-phosphate c2′-O-methylcytidine-5′-phosphate u 2′-O-methyluridine-5′-phosphate Ts2′-deoxy-thymidine-5′-phosphorothioate Q128 5′-(6-hydroxy hexyl)phosphate idT inverted 2′-deoxythymidine-3′-phosphate Abbeta-L-adenosine-3′-phosphate Ub beta-L-uridine-3′-phosphate Cbbeta-L-cytidine-3′-phosphate

The RNAi agents listed in Table 2 were prepared, as described in Example2.

Modified sequences were conceived for other 19-mers listed in Table 1.These modified sequences are listed in Table 3. As with Table 2, themodified sequences in Table 3 are examples and non-limiting; alternativemodifications are easily conceived by one of ordinary skill in the art,and additional or alternative base modifications are described elsewhereherein.

TABLE 3 MODIFIED SEQUENCES FOR ADDITIONAL 19-MERS SEQ ID SEQ ID PositionNO Sense 5′-3′modified NO Antisense 5′-3′ modified 201 1025GGGcccAGcAAcGucccGGdTdT 1537 CCGGGACGUUGCUGGGCCCdTdT 202 1026GGcccAGcAAcGucccGGcdTdT 1538 GCCGGGACGUUGCUGGGCCdTdT 203 1027GcccAGcAAcGucccGGccdTdT 1539 GGCCGGGACGUUGCUGGGCdTdT 204 1028cccAGcAAcGucccGGccudTdT 1540 AGGCCGGGACGUUGCUGGGdTdT 205 1029ccAGcAAcGucccGGccuudTdT 1541 AAGGCCGGGACGUUGCUGGdTdT 206 1030cAGcAAcGucccGGccuucdTdT 1542 GAAGGCCGGGACGUUGCUGdTdT 207 1031AGcAAcGucccGGccuuccdTdT 1543 GGAAGGCCGGGACGUUGCUdTdT 208 1032GcAAcGucccGGccuuccudTdT 1544 AGGAAGGCCGGGACGUUGCdTdT 248 1070GAGcGAcccGGAcAccGAcdTdT 1582 GUCGGUGUCCGGGUCGCUCdTdT 249 1071AGcGAcccGGAcAccGAcGdTdT 1583 CGUCGGUGUCCGGGUCGCUdTdT 250 1072GcGAcccGGAcAccGAcGcdTdT 1584 GCGUCGGUGUCCGGGUCGCdTdT 308 1077GuucGAccAGGGccAGuuudTdT 1589 AAACUGGCCCUGGUCGAACdTdT 437 1118GGucAAGccAGAGAGAGAcdTdT 1630 GUCUCUCUCUGGCUUGACCdTdT 438 1119GucAAGccAGAGAGAGAcGdTdT 1631 CGUCUCUCUCUGGCUUGACdTdT 439 1120ucAAGccAGAGAGAGAcGAdTdT 1632 UCGUCUCUCUCUGGCUUGAdTdT 440 1121cAAGccAGAGAGAGAcGAcdTdT 1633 GUCGUCUCUCUCUGGCUUGdTdT 441 1122AAGccAGAGAGAGAcGAcAdTdT 1634 UGUCGUCUCUCUCUGGCUUdTdT 442 1123AGccAGAGAGAGAcGAcAcdTdT 1635 GUGUCGUCUCUCUCUGGCUdTdT 443 1124GccAGAGAGAGAcGAcAcGdTdT 1636 CGUGUCGUCUCUCUCUGGCdTdT 444 1125ccAGAGAGAGAcGAcAcGGdTdT 1637 CCGUGUCGUCUCUCUCUGGdTdT 445 1126cAGAGAGAGAcGAcAcGGAdTdT 1638 UCCGUGUCGUCUCUCUCUGdTdT 446 1127AGAGAGAGAcGAcAcGGAGdTdT 1639 CUCCGUGUCGUCUCUCUCUdTdT 447 1128GAGAGAGAcGAcAcGGAGudTdT 1640 ACUCCGUGUCGUCUCUCUCdTdT 448 1129AGAGAGAcGAcAcGGAGuudTdT 1641 AACUCCGUGUCGUCUCUCUdTdT 449 1130GAGAGAcGAcAcGGAGuucdTdT 1642 GAACUCCGUGUCGUCUCUCdTdT 450 1131AGAGAcGAcAcGGAGuuccdTdT 1643 GGAACUCCGUGUCGUCUCUdTdT 451 1132GAGAcGAcAcGGAGuuccAdTdT 1644 UGGAACUCCGUGUCGUCUCdTdT 452 1133AGAcGAcAcGGAGuuccAGdTdT 1645 CUGGAACUCCGUGUCGUCUdTdT 453 1134GAcGAcAcGGAGuuccAGcdTdT 1646 GCUGGAACUCCGUGUCGUCdTdT 454 1135AcGAcAcGGAGuuccAGcAdTdT 1647 UGCUGGAACUCCGUGUCGUdTdT 455 1136cGAcAcGGAGuuccAGcAcdTdT 1648 GUGCUGGAACUCCGUGUCGdTdT 456 1137GAcAcGGAGuuccAGcAccdTdT 1649 GGUGCUGGAACUCCGUGUCdTdT 457 1138AcAcGGAGuuccAGcAcccdTdT 1650 GGGUGCUGGAACUCCGUGUdTdT 507 1157GAGAAcAucAAGAGGAAAGdTdT 1669 CUUUCCUCUUGAUGUUCUCdTdT 508 1158AGAAcAucAAGAGGAAAGudTdT 1670 ACUUUCCUCUUGAUGUUCUdTdT 686 1246GGccAGccuucGGcAGAAGdTdT 1758 CUUCUGCCGAAGGCUGGCCdTdT 687 1247GccAGccuucGGcAGAAGcdTdT 1759 GCUUCUGCCGAAGGCUGGCdTdT 688 1248ccAGccuucGGcAGAAGcAdTdT 1760 UGCUUCUGCCGAAGGCUGGdTdT 689 1249cAGccuucGGcAGAAGcAudTdT 1761 AUGCUUCUGCCGAAGGCUGdTdT 708 1268GcccAGcAAcAGAAAGucGdTdT 1780 CGACUUUCUGUUGCUGGGCdTdT 709 1269cccAGcAAcAGAAAGucGudTdT 1781 ACGACUUUCUGUUGCUGGGdTdT 710 1270ccAGcAAcAGAAAGucGucdTdT 1782 GACGACUUUCUGUUGCUGGdTdT 711 1271cAGcAAcAGAAAGucGucAdTdT 1783 UGACGACUUUCUGUUGCUGdTdT 712 1272AGcAAcAGAAAGucGucAAdTdT 1784 UUGACGACUUUCUGUUGCUdTdT 713 1273GcAAcAGAAAGucGucAAcdTdT 1785 GUUGACGACUUUCUGUUGCdTdT 714 1274cAAcAGAAAGucGucAAcAdTdT 1786 UGUUGACGACUUUCUGUUGdTdT 715 1275AAcAGAAAGucGucAAcAAdTdT 1787 UUGUUGACGACUUUCUGUUdTdT 716 1276AcAGAAAGucGucAAcAAGdTdT 1788 CUUGUUGACGACUUUCUGUdTdT 717 1277cAGAAAGucGucAAcAAGcdTdT 1789 GCUUGUUGACGACUUUCUGdTdT 718 1278AGAAAGucGucAAcAAGcudTdT 1790 AGCUUGUUGACGACUUUCUdTdT 719 1279GAAAGucGucAAcAAGcucdTdT 1791 GAGCUUGUUGACGACUUUCdTdT 720 1280AAAGucGucAAcAAGcucAdTdT 1792 UGAGCUUGUUGACGACUUUdTdT 721 1281AAGucGucAAcAAGcucAudTdT 1793 AUGAGCUUGUUGACGACUUdTdT 722 1282AGucGucAAcAAGcucAuudTdT 1794 AAUGAGCUUGUUGACGACUdTdT 723 1283GucGucAAcAAGcucAuucdTdT 1795 GAAUGAGCUUGUUGACGACdTdT 724 1284ucGucAAcAAGcucAuucAdTdT 1796 UGAAUGAGCUUGUUGACGAdTdT 725 1285cGucAAcAAGcucAuucAGdTdT 1797 CUGAAUGAGCUUGUUGACGdTdT 726 1286GucAAcAAGcucAuucAGudTdT 1798 ACUGAAUGAGCUUGUUGACdTdT 727 1287ucAAcAAGcucAuucAGuudTdT 1799 AACUGAAUGAGCUUGUUGAdTdT 728 1288cAAcAAGcucAuucAGuucdTdT 1800 GAACUGAAUGAGCUUGUUGdTdT 729 1289AAcAAGcucAuucAGuuccdTdT 1801 GGAACUGAAUGAGCUUGUUdTdT 730 1290AcAAGcucAuucAGuuccudTdT 1802 AGGAACUGAAUGAGCUUGUdTdT 863 1367GGAGcAcGuccAcGGcucGdTdT 1879 CGAGCCGUGGACGUGCUCCdTdT 864 1368GAGcAcGuccAcGGcucGGdTdT 1880 CCGAGCCGUGGACGUGCUCdTdT 865 1369AGcAcGuccAcGGcucGGGdTdT 1881 CCCGAGCCGUGGACGUGCUdTdT 866 1370GcAcGuccAcGGcucGGGcdTdT 1882 GCCCGAGCCGUGGACGUGCdTdT 867 1371cAcGuccAcGGcucGGGccdTdT 1883 GGCCCGAGCCGUGGACGUGdTdT 868 1372AcGuccAcGGcucGGGcccdTdT 1884 GGGCCCGAGCCGUGGACGUdTdT 954 1374GGAcccAucAucuccGAcAdTdT 1886 UGUCGGAGAUGAUGGGUCCdTdT 955 1375GAcccAucAucuccGAcAudTdT 1887 AUGUCGGAGAUGAUGGGUCdTdT 956 1376AcccAucAucuccGAcAucdTdT 1888 GAUGUCGGAGAUGAUGGGUdTdT 957 1377cccAucAucuccGAcAucAdTdT 1889 UGAUGUCGGAGAUGAUGGGdTdT 958 1378ccAucAucuccGAcAucAcdTdT 1890 GUGAUGUCGGAGAUGAUGGdTdT 959 1379cAucAucuccGAcAucAccdTdT 1891 GGUGAUGUCGGAGAUGAUGdTdT 960 1380AucAucuccGAcAucAccGdTdT 1892 CGGUGAUGUCGGAGAUGAUdTdT 961 1381ucAucuccGAcAucAccGAdTdT 1893 UCGGUGAUGUCGGAGAUGAdTdT 962 1382cAucuccGAcAucAccGAGdTdT 1894 CUCGGUGAUGUCGGAGAUGdTdT 963 1383AucuccGAcAucAccGAGcdTdT 1895 GCUCGGUGAUGUCGGAGAUdTdT 964 1384ucuccGAcAucAccGAGcudTdT 1896 AGCUCGGUGAUGUCGGAGAdTdT 1074 1414cccAGcccGccucAGAGccdTdT 1926 GGCUCUGAGGCGGGCUGGGdTdT 1075 1415ccAGcccGccucAGAGcccdTdT 1927 GGGCUCUGAGGCGGGCUGGdTdT 1376 1443GAGcAGccAcGGcuucAGcdTdT 1955 GCUGAAGCCGUGGCUGCUCdTdT 1377 1444AGcAGccAcGGcuucAGcGdTdT 1956 CGCUGAAGCCGUGGCUGCUdTdT 1378 1445GcAGccAcGGcuucAGcGudTdT 1957 ACGCUGAAGCCGUGGCUGCdTdT 1533 1501GAGAAcAGcAGcccGGAuudTdT 2013 AAUCCGGGCUGCUGUUCUCdTdT 1534 1502AGAAcAGcAGcccGGAuucdTdT 2014 GAAUCCGGGCUGCUGUUCUdTdT 1535 1503GAAcAGcAGcccGGAuucAdTdT 2015 UGAAUCCGGGCUGCUGUUCdTdT 1536 1504AAcAGcAGcccGGAuucAGdTdT 2016 CUGAAUCCGGGCUGCUGUUdTdT 1537 1505AcAGcAGcccGGAuucAGGdTdT 2017 CCUGAAUCCGGGCUGCUGUdTdT 1538 1506cAGcAGcccGGAuucAGGGdTdT 2018 CCCUGAAUCCGGGCUGCUGdTdT 1539 1507AGcAGcccGGAuucAGGGAdTdT 2019 UCCCUGAAUCCGGGCUGCUdTdT 1540 1508GcAGcccGGAuucAGGGAAdTdT 2020 UUCCCUGAAUCCGGGCUGCdTdT 1541 1509cAGcccGGAuucAGGGAAGdTdT 2021 CUUCCCUGAAUCCGGGCUGdTdT 1542 1510AGcccGGAuucAGGGAAGcdTdT 2022 GCUUCCCUGAAUCCGGGCUdTdT 1543 1511GcccGGAuucAGGGAAGcAdTdT 2023 UGCUUCCCUGAAUCCGGGCdTdT 1544 1512cccGGAuucAGGGAAGcAGdTdT 2024 CUGCUUCCCUGAAUCCGGGdTdT 1545 1513ccGGAuucAGGGAAGcAGcdTdT 2025 GCUGCUUCCCUGAAUCCGGdTdT 1546 1514cGGAuucAGGGAAGcAGcudTdT 2026 AGCUGCUUCCCUGAAUCCGdTdT 1725 1532GAGccucccAAAGccAAGGdTdT 3284 CCUUGGCUUUGGGAGGCUCdTdT 1726 1533AGccucccAAAGccAAGGAdTdT 3285 UCCUUGGCUUUGGGAGGCUdTdT 1727 1534GccucccAAAGccAAGGAcdTdT 3286 GUCCUUGGCUUUGGGAGGCdTdT 1728 1535ccucccAAAGccAAGGAccdTdT 3287 GGUCCUUGGCUUUGGGAGGdTdT 1729 1536cucccAAAGccAAGGAcccdTdT 3288 GGGUCCUUGGCUUUGGGAGdTdT

Example 1A Overlapping siRNAs

Some of the siRNAs listed above overlap each other in sequence. Thefollowing table presents a compilation of groups of RNAi agents, whereineach member of a group overlaps with each other member of the same groupby at least 12 nt. A 12-nt portion of the overlap of the sense strandsand a 12-nt portion of the overlap of the antisense strand arepresented. Thus, for example, AD-20296 and AD-20300 share the commontechnical feature of the sequence of CCAAGCUGUGGA in the sense strand,and the sequence of UCCACAGCUUGG in the antisense strand. Note of coursethat only a 12-nt portion of the overlap is shown; many groups of RNAiagents will overlap by more than 12 nt. The position within the gene isalso indicated.

TABLE 3A OVERLAPPING siRNAs RNAi Agents that SEQ SEQ ID overlap by atleast 12 nt Position Sense overlap ID NO: Antisense overlap NO: witheach other 210 ACGUCCCGGCCU 2097 AGGCCGGGACGU 2646 AD-20594, AD-20596211 CGUCCCGGCCUU 2098 AAGGCCGGGACG 2647 AD-20594, AD-20285 212GUCCCGGCCUUC 2099 GAAGGCCGGGAC 2648 AD-20594, AD-20285 213 UCCCGGCCUUCC2100 GGAAGGCCGGGA 2649 AD-20594, AD-20285 214 CCCGGCCUUCCU 2101AGGAAGGCCGGG 2650 AD-20594, AD-20598 215 CCGGCCUUCCUG 2102 CAGGAAGGCCGG2651 AD-20594, AD-20598 216 CGGCCUUCCUGA 2103 UCAGGAAGGCCG 2652AD-20598, AD-20288 217 GGCCUUCCUGAC 2104 GUCAGGAAGGCC 2653 AD-20289,AD-20598 218 GCCUUCCUGACC 2105 GGUCAGGAAGGC 2654 AD-20290, AD-20598 219CCUUCCUGACCA 2106 UGGUCAGGAAGG 2655 AD-20290, AD-20598 220 CUUCCUGACCAA2107 UUGGUCAGGAAG 2656 AD-20290, AD-20598 221 UUCCUGACCAAG 2108CUUGGUCAGGAA 2657 AD-20290, AD-20598 222 UCCUGACCAAGC 2109 GCUUGGUCAGGA2658 AD-20290, AD-20294 223 CCUGACCAAGCU 2110 AGCUUGGUCAGG 2659AD-20290, AD-20295 224 CUGACCAAGCUG 2111 CAGCUUGGUCAG 2660 AD-20290,AD-20296 225 UGACCAAGCUGU 2112 ACAGCUUGGUCA 2661 AD-20290, AD-20296 226GACCAAGCUGUG 2113 CACAGCUUGGUC 2662 AD-20297, AD-20296 227 ACCAAGCUGUGG2114 CCACAGCUUGGU 2663 AD-20296, AD-20299 228 CCAAGCUGUGGA 2115UCCACAGCUUGG 2664 AD-20300, AD-20296 229 CAAGCUGUGGAC 2116 GUCCACAGCUUG2665 AD-20300, AD-20296 230 AAGCUGUGGACC 2117 GGUCCACAGCUU 2666AD-20300, AD-20296 231 AGCUGUGGACCC 2118 GGGUCCACAGCU 2667 AD-20303,AD-20300, AD-20296 232 GCUGUGGACCCU 2119 AGGGUCCACAGC 2668 AD-20303,AD-20300 233 CUGUGGACCCUC 2120 GAGGGUCCACAG 2669 AD-20303, AD-20300 234UGUGGACCCUCG 2121 CGAGGGUCCACA 2670 AD-20303, AD-20300 235 GUGGACCCUCGU2122 ACGAGGGUCCAC 2671 AD-20303, AD-20300 236 UGGACCCUCGUG 2123CACGAGGGUCCA 2672 AD-20303, AD-20308 237 GGACCCUCGUGA 2124 UCACGAGGGUCC2673 AD-20303, AD-20308 238 GACCCUCGUGAG 2125 CUCACGAGGGUC 2674AD-20303, AD-20310 239 ACCCUCGUGAGC 2126 GCUCACGAGGGU 2675 AD-20310,AD-20311 240 CCCUCGUGAGCG 2127 CGCUCACGAGGG 2676 AD-20310, AD-20312 241CCUCGUGAGCGA 2128 UCGCUCACGAGG 2677 AD-20313, AD-20312 242 CUCGUGAGCGAC2129 GUCGCUCACGAG 2678 AD-20313, AD-20312 243 UCGUGAGCGACC 2130GGUCGCUCACGA 2679 AD-20315, AD-20313, AD-20312 244 CGUGAGCGACCC 2131GGGUCGCUCACG 2680 AD-20315, AD-20313, AD-20312 245 GUGAGCGACCCG 2132CGGGUCGCUCAC 2681 AD-20315, AD-20313, AD-20312 246 UGAGCGACCCGG 2133CCGGGUCGCUCA 2682 AD-20315, AD-20313, AD-20312 247 GAGCGACCCGGA 2134UCCGGGUCGCUC 2683 AD-20315, AD-20313, AD-20312 248 AGCGACCCGGAC 2135GUCCGGGUCGCU 2684 AD-20315, AD-20313 249 GCGACCCGGACA 2136 UGUCCGGGUCGC2685 AD-20315, AD-20318 250 CGACCCGGACAC 2137 GUGUCCGGGUCG 2686AD-20315, AD-20318 251 GACCCGGACACC 2138 GGUGUCCGGGUC 2687 AD-20318,AD-20317 252 ACCCGGACACCG 2139 CGGUGUCCGGGU 2688 AD-20318, AD-20317 270UCAUCUGCUGGA 2140 UCCAGCAGAUGA 2689 AD-20319, AD-20320 271 CAUCUGCUGGAG2141 CUCCAGCAGAUG 2690 AD-20319, AD-20320 272 AUCUGCUGGAGC 2142GCUCCAGCAGAU 2691 AD-20319, AD-20320 273 UCUGCUGGAGCC 2143 GGCUCCAGCAGA2692 AD-20319, AD-20320 274 CUGCUGGAGCCC 2144 GGGCUCCAGCAG 2693AD-20319, AD-20320 275 UGCUGGAGCCCG 2145 CGGGCUCCAGCA 2694 AD-20319,AD-20320 276 GCUGGAGCCCGA 2146 UCGGGCUCCAGC 2695 AD-20319, AD-20320 306UGUUCGACCAGG 2147 CCUGGUCGAACA 2696 AD-20345, AD-20344 307 GUUCGACCAGGG2148 CCCUGGUCGAAC 2697 AD-20345, AD-20344 308 UUCGACCAGGGC 2149GCCCUGGUCGAA 2698 AD-20345, AD-20344 309 UCGACCAGGGCC 2150 GGCCCUGGUCGA2699 AD-20345, AD-20344 310 CGACCAGGGCCA 2151 UGGCCCUGGUCG 2700AD-20345, AD-20348, AD-20344 311 GACCAGGGCCAG 2152 CUGGCCCUGGUC 2701AD-20349, AD-20345, AD-20348, AD-20344 312 ACCAGGGCCAGU 2153ACUGGCCCUGGU 2702 AD-20349, AD-20345, AD-20348, AD-20344 313CCAGGGCCAGUU 2154 AACUGGCCCUGG 2703 AD-20349, AD-20345, AD-20348 314CAGGGCCAGUUU 2155 AAACUGGCCCUG 2704 AD-20349, AD-20348 315 AGGGCCAGUUUG2156 CAAACUGGCCCU 2705 AD-20349, AD-20353, AD-20348 316 GGGCCAGUUUGC2157 GCAAACUGGCCC 2706 AD-20349, AD-20353, AD-20348 317 GGCCAGUUUGCC2158 GGCAAACUGGCC 2707 AD-20349, AD-20353, AD-20348 318 GCCAGUUUGCCA2159 UGGCAAACUGGC 2708 AD-20349, AD-20353 319 CCAGUUUGCCAA 2160UUGGCAAACUGG 2709 AD-20353, AD-20356 320 CAGUUUGCCAAG 2161 CUUGGCAAACUG2710 AD-20353, AD-20358 321 AGUUUGCCAAGG 2162 CCUUGGCAAACU 2711AD-20353, AD-20359 322 GUUUGCCAAGGA 2163 UCCUUGGCAAAC 2712 AD-20353,AD-20360 323 UUUGCCAAGGAG 2164 CUCCUUGGCAAA 2713 AD-20361, AD-20360 324UUGCCAAGGAGG 2165 CCUCCUUGGCAA 2714 AD-20362, AD-20360 325 UGCCAAGGAGGU2166 ACCUCCUUGGCA 2715 AD-20362, AD-20360 326 GCCAAGGAGGUG 2167CACCUCCUUGGC 2716 AD-20364, AD-20362 327 CCAAGGAGGUGC 2168 GCACCUCCUUGG2717 AD-20365, AD-20364, AD-20362 328 CAAGGAGGUGCU 2169 AGCACCUCCUUG2718 AD-20365, AD-20366, AD-20364, AD-20362 329 AAGGAGGUGCUG 2170CAGCACCUCCUU 2719 AD-20365, AD-20366, AD-20364, AD-20362 330AGGAGGUGCUGC 2171 GCAGCACCUCCU 2720 AD-20365, AD-20366, AD-20364,AD-20362 331 GGAGGUGCUGCC 2172 GGCAGCACCUCC 2721 AD-20365, AD-20366,AD-20364, AD-20362 332 GAGGUGCUGCCC 2173 GGGCAGCACCUC 2722 AD-20365,AD-20366, AD-20364 333 AGGUGCUGCCCA 2174 UGGGCAGCACCU 2723 AD-20365,AD-20366, AD-20364 334 GGUGCUGCCCAA 2175 UUGGGCAGCACC 2724 AD-20365,AD-20366 335 GUGCUGCCCAAG 2176 CUUGGGCAGCAC 2725 AD-20366, AD-20367 336UGCUGCCCAAGU 2177 ACUUGGGCAGCA 2726 AD-20368, AD-20367 341 CCCAAGUACUUC2178 GAAGUACUUGGG 2727 AD-20282, AD-20281 342 CCAAGUACUUCA 2179UGAAGUACUUGG 2728 AD-20282, AD-20281 343 CAAGUACUUCAA 2180 UUGAAGUACUUG2729 AD-20282, AD-20281 344 AAGUACUUCAAG 2181 CUUGAAGUACUU 2730AD-20282, AD-20281 345 AGUACUUCAAGC 2182 GCUUGAAGUACU 2731 AD-20282,AD-20281 346 GUACUUCAAGCA 2183 UGCUUGAAGUAC 2732 AD-20282, AD-20281 347UACUUCAAGCAC 2184 GUGCUUGAAGUA 2733 AD-20282, AD-20281 348 ACUUCAAGCACA2185 UGUGCUUGAAGU 2734 AD-20282, AD-20283 351 UCAAGCACAACA 2186UGUUGUGCUUGA 2735 AD-20369, AD-20370 352 CAAGCACAACAA 2187 UUGUUGUGCUUG2736 AD-20369, AD-20370 353 AAGCACAACAAC 2188 GUUGUUGUGCUU 2737AD-20369, AD-20370 354 AGCACAACAACA 2189 UGUUGUUGUGCU 2738 AD-20373,AD-20369 355 GCACAACAACAU 2190 AUGUUGUUGUGC 2739 AD-20373, AD-20374 356CACAACAACAUG 2191 CAUGUUGUUGUG 2740 AD-20373, AD-20374 357 ACAACAACAUGG2192 CCAUGUUGUUGU 2741 AD-20373, AD-20374, AD-20376 358 CAACAACAUGGC2193 GCCAUGUUGUUG 2742 AD-20373, AD-20376 359 AACAACAUGGCC 2194GGCCAUGUUGUU 2743 AD-20373, AD-20374, AD-20376, AD-20378 360ACAACAUGGCCA 2195 UGGCCAUGUUGU 2744 AD-20373, AD-20374, AD-20376,AD-20378 361 CAACAUGGCCAG 2196 CUGGCCAUGUUG 2745 AD-20373, AD-20374,AD-20376, AD-20378, AD-20380 362 AACAUGGCCAGC 2197 GCUGGCCAUGUU 2746AD-20374, AD-20376, AD-20378, AD-20380 363 ACAUGGCCAGCU 2198AGCUGGCCAUGU 2747 AD-20376, AD-20378, AD-20380 364 CAUGGCCAGCUU 2199AAGCUGGCCAUG 2748 AD-20376, AD-20378, AD-20380 365 AUGGCCAGCUUC 2200GAAGCUGGCCAU 2749 AD-20378, AD-20380 366 UGGCCAGCUUCG 2201 CGAAGCUGGCCA2750 AD-20378, AD-20380 367 GGCCAGCUUCGU 2202 ACGAAGCUGGCC 2751AD-20380, AD-20385 368 GCCAGCUUCGUG 2203 CACGAAGCUGGC 2752 AD-20380,AD-20385 369 CCAGCUUCGUGC 2204 GCACGAAGCUGG 2753 AD-20382, AD-20385 370CAGCUUCGUGCG 2205 CGCACGAAGCUG 2754 AD-20382, AD-20385 371 AGCUUCGUGCGG2206 CCGCACGAAGCU 2755 AD-20384, AD-20385 372 GCUUCGUGCGGC 2207GCCGCACGAAGC 2756 AD-20384, AD-20385 489 GCCAGGAGCAGC 2208 GCUGCUCCUGGC2757 AD-20388, AD-20387 490 CCAGGAGCAGCU 2209 AGCUGCUCCUGG 2758AD-20388, AD-20387, AD-20389 491 CAGGAGCAGCUC 2210 GAGCUGCUCCUG 2759AD-20390, AD-20388, AD-20387, AD-20389 492 AGGAGCAGCUCC 2211GGAGCUGCUCCU 2760 AD-20390, AD-20391, AD-20388, AD-20387, AD-20389 493GGAGCAGCUCCU 2212 AGGAGCUGCUCC 2761 AD-20390, AD-20391, AD-20388,AD-20392, AD-20387, AD-20389 494 GAGCAGCUCCUU 2213 AAGGAGCUGCUC 2762AD-20390, AD-20391, AD-20388, AD-20393, AD-20392, AD-20387, AD-20389 495AGCAGCUCCUUG 2214 CAAGGAGCUGCU 2763 AD-20390, AD-20391, AD-20388,AD-20393, AD-20392, AD-20387, AD-20389 496 GCAGCUCCUUGA 2215UCAAGGAGCUGC 2764 AD-20390, AD-20395, AD-20391, AD-20388, AD-20393,AD-20392, AD-20389 497 CAGCUCCUUGAG 2216 CUCAAGGAGCUG 2765 AD-20390,AD-20395, AD-20396, AD-20391, AD-20393, AD-20392, AD-20389 498AGCUCCUUGAGA 2217 UCUCAAGGAGCU 2766 AD-20390, AD-20395, AD-20396,AD-20397, AD-20391, AD-20393, AD-20392 499 GCUCCUUGAGAA 2218UUCUCAAGGAGC 2767 AD-20395, AD-20396, AD-20398, AD-20397, AD-20391,AD-20393, AD-20392 500 CUCCUUGAGAAC 2219 GUUCUCAAGGAG 2768 AD-20395,AD-20396, AD-20398, AD-20397, AD-20393, AD-20399, AD-20392 501UCCUUGAGAACA 2220 UGUUCUCAAGGA 2769 AD-20395, AD-20396, AD-20398,AD-20397, AD-20393, AD-20399 502 CCUUGAGAACAU 2221 AUGUUCUCAAGG 2770AD-20395, AD-20396, AD-20398, AD-20397, AD-20399, AD-20401 503CUUGAGAACAUC 2222 GAUGUUCUCAAG 2771 AD-20395, AD-20396, AD-20402,AD-20398, AD-20397, AD-20399, AD-20401 504 UUGAGAACAUCA 2223UGAUGUUCUCAA 2772 AD-20403, AD-20396, AD-20402, AD-20398, AD-20397,AD-20399, AD-20401 505 UGAGAACAUCAA 2224 UUGAUGUUCUCA 2773 AD-20403,AD-20404, AD-20402, AD-20398, AD-20397, AD-20399, AD-20401 506GAGAACAUCAAG 2225 CUUGAUGUUCUC 2774 AD-20403, AD-20404, AD-20402,AD-20398, AD-20399, AD-20401 507 AGAACAUCAAGA 2226 UCUUGAUGUUCU 2775AD-20403, AD-20404, AD-20402, AD-20399, AD-20401 508 GAACAUCAAGAG 2227CUCUUGAUGUUC 2776 AD-20403, AD-20404, AD-20402, AD-20401 509AACAUCAAGAGG 2228 CCUCUUGAUGUU 2777 AD-20403, AD-20404, AD-20402,AD-20406, AD-20401 510 ACAUCAAGAGGA 2229 UCCUCUUGAUGU 2778 AD-20403,AD-20404, AD-20402, AD-20406, AD-20407 511 CAUCAAGAGGAA 2230UUCCUCUUGAUG 2779 AD-20403, AD-20404, AD-20406, AD-20407, AD-20408 512AUCAAGAGGAAA 2231 UUUCCUCUUGAU 2780 AD-20409, AD-20404, AD-20406,AD-20407, AD-20408 513 UCAAGAGGAAAG 2232 CUUUCCUCUUGA 2781 AD-20409,AD-20406, AD-20410, AD-20407, AD-20408 514 CAAGAGGAAAGU 2233ACUUUCCUCUUG 2782 AD-20409, AD-20406, AD-20410, AD-20407, AD-20408,AD-20411 515 AAGAGGAAAGUG 2234 CACUUUCCUCUU 2783 AD-20409, AD-20406,AD-20410, AD-20407, AD-20408, AD-20411 516 AGAGGAAAGUGA 2235UCACUUUCCUCU 2784 AD-20409, AD-20406, AD-20410, AD-20407, AD-20408,AD-20413, AD-20411 517 GAGGAAAGUGAC 2236 GUCACUUUCCUC 2785 AD-20409,AD-20410, AD-20407, AD-20408, AD-20413, AD-20411 518 AGGAAAGUGACC 2237GGUCACUUUCCU 2786 AD-20409, AD-20410, AD-20408, AD-20413, AD-20411 519GGAAAGUGACCA 2238 UGGUCACUUUCC 2787 AD-20409, AD-20410, AD-20413,AD-20411 520 GAAAGUGACCAG 2239 CUGGUCACUUUC 2788 AD-20410, AD-20413,AD-20411 521 AAAGUGACCAGU 2240 ACUGGUCACUUU 2789 AD-20413, AD-20411 522AAGUGACCAGUG 2241 CACUGGUCACUU 2790 AD-20413, AD-20412 523 AGUGACCAGUGU2242 ACACUGGUCACU 2791 AD-20420, AD-20413 524 GUGACCAGUGUG 2243CACACUGGUCAC 2792 AD-20420, AD-20421 525 UGACCAGUGUGU 2244 ACACACUGGUCA2793 AD-20421, AD-20422 526 GACCAGUGUGUC 2245 GACACACUGGUC 2794AD-20421, AD-20422 527 ACCAGUGUGUCC 2246 GGACACACUGGU 2795 AD-20421,AD-20422 528 CCAGUGUGUCCA 2247 UGGACACACUGG 2796 AD-20421, AD-20422 529CAGUGUGUCCAC 2248 GUGGACACACUG 2797 AD-20426, AD-20421, AD-20422, 530AGUGUGUCCACC 2249 GGUGGACACACU 2798 AD-20427, AD-20426, AD-20421,AD-20422 531 GUGUGUCCACCC 2250 GGGUGGACACAC 2799 AD-20427, AD-20426,AD-20421, AD-20422, AD-20428 532 UGUGUCCACCCU 2251 AGGGUGGACACA 2800AD-20427, AD-20426, AD-20422, AD-20428 533 GUGUCCACCCUG 2252CAGGGUGGACAC 2801 AD-20427, AD-20426, AD-20428 534 UGUCCACCCUGA 2253UCAGGGUGGACA 2802 AD-20427, AD-20426, AD-20428 535 GUCCACCCUGAA 2254UUCAGGGUGGAC 2803 AD-20427, AD-20426, AD-20428 536 UCCACCCUGAAG 2255CUUCAGGGUGGA 2804 AD-20427, AD-20426, AD-20433, AD-20428 537CCACCCUGAAGA 2256 UCUUCAGGGUGG 2805 AD-20434, AD-20427, AD-20433,AD-20428 538 CACCCUGAAGAG 2257 CUCUUCAGGGUG 2806 AD-20434, AD-20433,AD-20428, AD-20435 539 ACCCUGAAGAGU 2258 ACUCUUCAGGGU 2807 AD-20434,AD-20433, AD-20436, AD-20435 540 CCCUGAAGAGUG 2259 CACUCUUCAGGG 2808AD-20434, AD-20433, AD-20436, AD-20437, AD-20435 541 CCUGAAGAGUGA 2260UCACUCUUCAGG 2809 AD-20434, AD-20438, AD-20433, AD-20436, AD-20437,AD-20435 542 CUGAAGAGUGAA 2261 UUCACUCUUCAG 2810 AD-20439, AD-20434,AD-20438, AD-20433, AD-20436, AD-20437, AD-20435 543 UGAAGAGUGAAG 2262CUUCACUCUUCA 2811 AD-20439, AD-20434, AD-20487, AD-20438, AD-20433,AD-20436, AD-20437, AD-20435 544 GAAGAGUGAAGA 2263 UCUUCACUCUUC 2812AD-20439, AD-20434, AD-20487, AD-20488, AD-20438, AD-20436, AD-20437,AD-20435 545 AAGAGUGAAGAC 2264 GUCUUCACUCUU 2813 AD-20439, AD-20487,AD-20488, AD-20489, AD-20438, AD-20436, AD-20437, AD-20435 546AGAGUGAAGACA 2265 UGUCUUCACUCU 2814 AD-20439, AD-20487, AD-20488,AD-20489, AD-20438, AD-20436, AD-20437, AD-20490 547 GAGUGAAGACAU 2266AUGUCUUCACUC 2815 AD-20439, AD-20491, AD-20487, AD-20488, AD-20489,AD-20438, AD-20437, AD-20490 548 AGUGAAGACAUA 2267 UAUGUCUUCACU 2816AD-20439, AD-20491, AD-20487, AD-20488, AD-20489, AD-20438, AD-20490 549GUGAAGACAUAA 2268 UUAUGUCUUCAC 2817 AD-20439, AD-20491, AD-20487,AD-20488, AD-20489, AD-20493, AD-20490 550 UGAAGACAUAAA 2269UUUAUGUCUUCA 2818 AD-20491, AD-20487, AD-20488, AD-20489, AD-20493,AD-20490 551 GAAGACAUAAAG 2270 CUUUAUGUCUUC 2819 AD-20491, AD-20488,AD-20489, AD-20493, AD-20490 552 AAGACAUAAAGA 2271 UCUUUAUGUCUU 2820AD-20491, AD-20489, AD-20493, AD-20490 553 AGACAUAAAGAU 2272AUCUUUAUGUCU 2821 AD-20491, AD-20493, AD-20490 554 GACAUAAAGAUC 2273GAUCUUUAUGUC 2822 AD-20491, AD-20493 555 ACAUAAAGAUCC 2274 GGAUCUUUAUGU2823 AD-20492, AD-20493 579 UCACCAAGCUGC 2275 GCAGCUUGGUGA 2824AD-20494, AD-20495 580 CACCAAGCUGCU 2276 AGCAGCUUGGUG 2825 AD-20494,AD-20495 581 ACCAAGCUGCUG 2277 CAGCAGCUUGGU 2826 AD-20494, AD-20495 582CCAAGCUGCUGA 2278 UCAGCAGCUUGG 2827 AD-20494, AD-20495 583 CAAGCUGCUGAC2279 GUCAGCAGCUUG 2828 AD-20494, AD-20495 584 AAGCUGCUGACG 2280CGUCAGCAGCUU 2829 AD-20494, AD-20495 585 AGCUGCUGACGG 2281 CCGUCAGCAGCU2830 AD-20494, AD-20495, AD-20501 586 GCUGCUGACGGA 2282 UCCGUCAGCAGC2831 AD-20495, AD-20502, AD-20501 587 CUGCUGACGGAC 2283 GUCCGUCAGCAG2832 AD-20502, AD-20501 588 UGCUGACGGACG 2284 CGUCCGUCAGCA 2833AD-20504, AD-20502, AD-20501 589 GCUGACGGACGU 2285 ACGUCCGUCAGC 2834AD-20504, AD-20502, AD-20501 590 CUGACGGACGUG 2286 CACGUCCGUCAG 2835AD-20504, AD-20506, AD-20502, AD-20501 591 UGACGGACGUGC 2287GCACGUCCGUCA 2836 AD-20504, AD-20506, AD-20507, AD-20502, AD-20501 592GACGGACGUGCA 2288 UGCACGUCCGUC 2837 AD-20504, AD-20506, AD-20507,AD-20502, AD-20501 593 ACGGACGUGCAG 2289 CUGCACGUCCGU 2838 AD-20504,AD-20506, AD-20507, AD-20502 594 CGGACGUGCAGC 2290 GCUGCACGUCCG 2839AD-20504, AD-20506, AD-20507, AD-20510 595 GGACGUGCAGCU 2291AGCUGCACGUCC 2840 AD-20511, AD-20504, AD-20506, AD-20507, AD-20510 596GACGUGCAGCUG 2292 CAGCUGCACGUC 2841 AD-20511, AD-20506, AD-20507,AD-20510 597 ACGUGCAGCUGA 2293 UCAGCUGCACGU 2842 AD-20511, AD-20506,AD-20507, AD-20510, AD-20513 598 CGUGCAGCUGAU 2294 AUCAGCUGCACG 2843AD-20511, AD-20507, AD-20510, AD-20513 599 GUGCAGCUGAUG 2295CAUCAGCUGCAC 2844 AD-20511, AD-20510, AD-20513 600 UGCAGCUGAUGA 2296UCAUCAGCUGCA 2845 AD-20511, AD-20510, AD-20513 601 GCAGCUGAUGAA 2297UUCAUCAGCUGC 2846 AD-20511, AD-20510, AD-20513 602 CAGCUGAUGAAG 2298CUUCAUCAGCUG 2847 AD-20511, AD-20513 603 AGCUGAUGAAGG 2299 CCUUCAUCAGCU2848 AD-20512, AD-20513 660 AGAAUGAGGCUC 2300 GAGCCUCAUUCU 2849AD-20514, AD-20515 661 GAAUGAGGCUCU 2301 AGAGCCUCAUUC 2850 AD-20516,AD-20515 662 AAUGAGGCUCUG 2302 CAGAGCCUCAUU 2851 AD-20517, AD-20515 663AUGAGGCUCUGU 2303 ACAGAGCCUCAU 2852 AD-20518, AD-20515 664 UGAGGCUCUGUG2304 CACAGAGCCUCA 2853 AD-20519, AD-20515 665 GAGGCUCUGUGG 2305CCACAGAGCCUC 2854 AD-20520, AD-20515 666 AGGCUCUGUGGC 2306 GCCACAGAGCCU2855 AD-20521, AD-20515 667 GGCUCUGUGGCG 2307 CGCCACAGAGCC 2856AD-20521, AD-20522 668 GCUCUGUGGCGG 2308 CCGCCACAGAGC 2857 AD-20523,AD-20522 669 CUCUGUGGCGGG 2309 CCCGCCACAGAG 2858 AD-20524, AD-20523, 670UCUGUGGCGGGA 2310 UCCCGCCACAGA 2859 AD-20524, AD-20525, 671 CUGUGGCGGGAG2311 CUCCCGCCACAG 2860 AD-20524, AD-20525 672 UGUGGCGGGAGG 2312CCUCCCGCCACA 2861 AD-20527, AD-20524 673 GUGGCGGGAGGU 2313 ACCUCCCGCCAC2862 AD-20527, AD-20524 674 UGGCGGGAGGUG 2314 CACCUCCCGCCA 2863AD-20527, AD-20524 675 GGCGGGAGGUGG 2315 CCACCUCCCGCC 2864 AD-20530,AD-20527 676 GCGGGAGGUGGC 2316 GCCACCUCCCGC 2865 AD-20530, AD-20527 677CGGGAGGUGGCC 2317 GGCCACCUCCCG 2866 AD-20530, AD-20527 678 GGGAGGUGGCCA2318 UGGCCACCUCCC 2867 AD-20530, AD-20527 679 GGAGGUGGCCAG 2319CUGGCCACCUCC 2868 AD-20530, AD-20527 680 GAGGUGGCCAGC 2320 GCUGGCCACCUC2869 AD-20530, AD-20535 681 AGGUGGCCAGCC 2321 GGCUGGCCACCU 2870AD-20530, AD-20535 682 GGUGGCCAGCCU 2322 AGGCUGGCCACC 2871 AD-20530,AD-20535 683 GUGGCCAGCCUU 2323 AAGGCUGGCCAC 2872 AD-20532, AD-20535 684UGGCCAGCCUUC 2324 GAAGGCUGGCCA 2873 AD-20539, AD-20535 685 GGCCAGCCUUCG2325 CGAAGGCUGGCC 2874 AD-20539, AD-20535 686 GCCAGCCUUCGG 2326CCGAAGGCUGGC 2875 AD-20539, AD-20535 687 CCAGCCUUCGGC 2327 GCCGAAGGCUGG2876 AD-20539, AD-20535 688 CAGCCUUCGGCA 2328 UGCCGAAGGCUG 2877AD-20539, AD-20538 689 AGCCUUCGGCAG 2329 CUGCCGAAGGCU 2878 AD-20539,AD-20540 690 GCCUUCGGCAGA 2330 UCUGCCGAAGGC 2879 AD-20539, AD-20540 691CCUUCGGCAGAA 2331 UUCUGCCGAAGG 2880 AD-20539, AD-20540 692 CUUCGGCAGAAG2332 CUUCUGCCGAAG 2881 AD-20541, AD-20540 693 UUCGGCAGAAGC 2333GCUUCUGCCGAA 2882 AD-20544, AD-20540 694 UCGGCAGAAGCA 2334 UGCUUCUGCCGA2883 AD-20544, AD-20545 695 CGGCAGAAGCAU 2335 AUGCUUCUGCCG 2884AD-20546, AD-20544, AD-20545 696 GGCAGAAGCAUG 2336 CAUGCUUCUGCC 2885AD-20546, AD-20544, AD-20545, AD-20547 697 GCAGAAGCAUGC 2337GCAUGCUUCUGC 2886 AD-20546, AD-20544, AD-20545, AD-20548, AD-20547 698CAGAAGCAUGCC 2338 GGCAUGCUUCUG 2887 AD-20546, AD-20544, AD-20545,AD-20549, AD-20548, AD-20547 699 AGAAGCAUGCCC 2339 GGGCAUGCUUCU 2888AD-20546, AD-20544, AD-20545, AD-20549, AD-20548, AD-20547 700GAAGCAUGCCCA 2340 UGGGCAUGCUUC 2889 AD-20546, AD-20544, AD-20545,AD-20549, AD-20548, AD-20547 701 AAGCAUGCCCAG 2341 CUGGGCAUGCUU 2890AD-20546, AD-20552, AD-20545, AD-20549, AD-20548, AD-20547 702AGCAUGCCCAGC 2342 GCUGGGCAUGCU 2891 AD-20546, AD-20552, AD-20549,AD-20548, AD-20547 703 GCAUGCCCAGCA 2343 UGCUGGGCAUGC 2892 AD-20552,AD-20549, AD-20548, AD-20547 704 CAUGCCCAGCAA 2344 UUGCUGGGCAUG 2893AD-20552, AD-20555, AD-20549, AD-20548 705 AUGCCCAGCAAC 2345GUUGCUGGGCAU 2894 AD-20556, AD-20552, AD-20555, AD-20549 706UGCCCAGCAACA 2346 UGUUGCUGGGCA 2895 AD-20557, AD-20556, AD-20552,AD-20555 707 GCCCAGCAACAG 2347 CUGUUGCUGGGC 2896 AD-20557, AD-20556,AD-20552, AD-20555 708 CCCAGCAACAGA 2348 UCUGUUGCUGGG 2897 AD-20557,AD-20556, AD-20552, AD-20555 709 CCAGCAACAGAA 2349 UUCUGUUGCUGG 2898AD-20557, AD-20556, AD-20555 710 CAGCAACAGAAA 2350 UUUCUGUUGCUG 2899AD-20557, AD-20556, AD-20555 711 AGCAACAGAAAG 2351 CUUUCUGUUGCU 2900AD-20557, AD-20556, AD-20555 712 GCAACAGAAAGU 2352 ACUUUCUGUUGC 2901AD-20557, AD-20556 731 AAGCUCAUUCAG 2353 CUGAAUGAGCUU 2902 AD-20559,AD-20558 732 AGCUCAUUCAGU 2354 ACUGAAUGAGCU 2903 AD-20559, AD-20558,AD-20560 733 GCUCAUUCAGUU 2355 AACUGAAUGAGC 2904 AD-20559, AD-20558,AD-20560, AD-20561 734 CUCAUUCAGUUC 2356 GAACUGAAUGAG 2905 AD-20562,AD-20559, AD-20558, AD-20560, AD-20561 735 UCAUUCAGUUCC 2357GGAACUGAAUGA 2906 AD-20562, AD-20559, AD-20563, AD-20558, AD-20560,AD-20561 736 CAUUCAGUUCCU 2358 AGGAACUGAAUG 2907 AD-20562, AD-20559,AD-20563, AD-20558, AD-20560, AD-20564, AD-20561 737 AUUCAGUUCCUG 2359CAGGAACUGAAU 2908 AD-20562, AD-20559, AD-20563, AD-20558, AD-20560,AD-20564, AD-20565, AD-20561 738 UUCAGUUCCUGA 2360 UCAGGAACUGAA 2909AD-20566, AD-20562, AD-20559, AD-20563, AD-20560, AD-20564, AD-20565,AD-20561 739 UCAGUUCCUGAU 2361 AUCAGGAACUGA 2910 AD-20566, AD-20562,AD-20563, AD-20560, AD-20564, AD-20565, AD-20561 740 CAGUUCCUGAUC 2362GAUCAGGAACUG 2911 AD-20566, AD-20562, AD-20563, AD-20564, AD-20565,AD-20561 741 AGUUCCUGAUCU 2363 AGAUCAGGAACU 2912 AD-20569, AD-20566,AD-20562, AD-20563, AD-20564, AD-20565 742 GUUCCUGAUCUC 2364GAGAUCAGGAAC 2913 AD-20569, AD-20566, AD-20563, AD-20570, AD-20564,AD-20565 743 UUCCUGAUCUCA 2365 UGAGAUCAGGAA 2914 AD-20569, AD-20566,AD-20570, AD-20564, AD-20565 744 UCCUGAUCUCAC 2366 GUGAGAUCAGGA 2915AD-20569, AD-20566, AD-20572, AD-20570, AD-20565 745 CCUGAUCUCACU 2367AGUGAGAUCAGG 2916 AD-20569, AD-20566, AD-20572, AD-20570 746CUGAUCUCACUG 2368 CAGUGAGAUCAG 2917 AD-20569, AD-20572, AD-20570,AD-20574 747 UGAUCUCACUGG 2369 CCAGUGAGAUCA 2918 AD-20569, AD-20572,AD-20570, AD-20574, AD-20575 748 GAUCUCACUGGU 2370 ACCAGUGAGAUC 2919AD-20569, AD-20576, AD-20572, AD-20570, AD-20574, AD-20575 749AUCUCACUGGUG 2371 CACCAGUGAGAU 2920 AD-20576, AD-20577, AD-20572,AD-20570, AD-20574, AD-20575 750 UCUCACUGGUGC 2372 GCACCAGUGAGA 2921AD-20578, AD-20576, AD-20577, AD-20572, AD-20574, AD-20575 751CUCACUGGUGCA 2373 UGCACCAGUGAG 2922 AD-20578, AD-20576, AD-20579,AD-20577, AD-20572, AD-20574, AD-20575 752 UCACUGGUGCAG 2374CUGCACCAGUGA 2923 AD-20580, AD-20578, AD-20576, AD-20579, AD-20577,AD-20574, AD-20575 753 CACUGGUGCAGU 2375 ACUGCACCAGUG 2924 AD-20580,AD-20578, AD-20576, AD-20579, AD-20577, AD-20574, AD-20575 754ACUGGUGCAGUC 2376 GACUGCACCAGU 2925 AD-20580, AD-20578, AD-20576,AD-20579, AD-20577, AD-20582, AD-20575 755 CUGGUGCAGUCA 2377UGACUGCACCAG 2926 AD-20580, AD-20578, AD-20576, AD-20579, AD-20577,AD-20582, AD-20625 756 UGGUGCAGUCAA 2378 UUGACUGCACCA 2927 AD-20580,AD-20626, AD-20578, AD-20579, AD-20577, AD-20582, AD-20625 757GGUGCAGUCAAA 2379 UUUGACUGCACC 2928 AD-20580, AD-20627, AD-20626,AD-20578, AD-20579, AD-20582, AD-20625 758 GUGCAGUCAAAC 2380GUUUGACUGCAC 2929 AD-20580, AD-20627, AD-20626, AD-20579, AD-20582,AD-20625 759 UGCAGUCAAACC 2381 GGUUUGACUGCA 2930 AD-20580, AD-20627,AD-20626, AD-20582, AD-20629, AD-20625 760 GCAGUCAAACCG 2382CGGUUUGACUGC 2931 AD-20627, AD-20626, AD-20582, AD-20629, AD-20625 761CAGUCAAACCGG 2383 CCGGUUUGACUG 2932 AD-20627, AD-20626, AD-20582,AD-20629, AD-20625 762 AGUCAAACCGGA 2384 UCCGGUUUGACU 2933 AD-20627,AD-20626, AD-20629, AD-20625 763 GUCAAACCGGAU 2385 AUCCGGUUUGAC 2934AD-20627, AD-20626, AD-20629 764 UCAAACCGGAUC 2386 GAUCCGGUUUGA 2935AD-20627, AD-20629 765 CAAACCGGAUCC 2387 GGAUCCGGUUUG 2936 AD-20628,AD-20629 766 AAACCGGAUCCU 2388 AGGAUCCGGUUU 2937 AD-20630, AD-20629 767AACCGGAUCCUG 2389 CAGGAUCCGGUU 2938 AD-20630, AD-20632 768 ACCGGAUCCUGG2390 CCAGGAUCCGGU 2939 AD-20632, AD-20631 799 CCUGAUGCUGAA 2391UUCAGCAUCAGG 2940 AD-20634, AD-20635 800 CUGAUGCUGAAC 2392 GUUCAGCAUCAG2941 AD-20634, AD-20636 801 UGAUGCUGAACG 2393 CGUUCAGCAUCA 2942AD-20637, AD-20634 802 GAUGCUGAACGA 2394 UCGUUCAGCAUC 2943 AD-20634,AD-20638 803 AUGCUGAACGAC 2395 GUCGUUCAGCAU 2944 AD-20639, AD-20634,AD-20638 804 UGCUGAACGACA 2396 UGUCGUUCAGCA 2945 AD-20640, AD-20639,AD-20634, AD-20638 805 GCUGAACGACAG 2397 CUGUCGUUCAGC 2946 AD-20641,AD-20640, AD-20639, AD-20634, AD-20638 806 CUGAACGACAGU 2398ACUGUCGUUCAG 2947 AD-20641, AD-20640, AD-20639, AD-20638 807UGAACGACAGUG 2399 CACUGUCGUUCA 2948 AD-20641, AD-20643, AD-20640,AD-20639, AD-20638 808 GAACGACAGUGG 2400 CCACUGUCGUUC 2949 AD-20641,AD-20643, AD-20640, AD-20639, AD-20638, AD-20644 809 AACGACAGUGGC 2401GCCACUGUCGUU 2950 AD-20641, AD-20643, AD-20640, AD-20639, AD-20638,AD-20644 810 ACGACAGUGGCU 2402 AGCCACUGUCGU 2951 AD-20646, AD-20643,AD-20640, AD-20639, AD-20644 811 CGACAGUGGCUC 2403 GAGCCACUGUCG 2952AD-20646, AD-20643, AD-20640, AD-20644 812 GACAGUGGCUCA 2404UGAGCCACUGUC 2953 AD-20646, AD-20643, AD-20648, AD-20644 813ACAGUGGCUCAG 2405 CUGAGCCACUGU 2954 AD-20646, AD-20643, AD-20648,AD-20644 814 CAGUGGCUCAGC 2406 GCUGAGCCACUG 2955 AD-20646, AD-20643,AD-20650, AD-20648, AD-20644 815 AGUGGCUCAGCA 2407 UGCUGAGCCACU 2956AD-20651, AD-20646, AD-20650, AD-20648, AD-20644 816 GUGGCUCAGCAC 2408GUGCUGAGCCAC 2957 AD-20651, AD-20646, AD-20650, AD-20652, AD-20648 817UGGCUCAGCACA 2409 UGUGCUGAGCCA 2958 AD-20651, AD-20646, AD-20653,AD-20650, AD-20652, AD-20648 818 GGCUCAGCACAU 2410 AUGUGCUGAGCC 2959AD-20653, AD-20650, AD-20652, AD-20648 819 GCUCAGCACAUU 2411AAUGUGCUGAGC 2960 AD-20653, AD-20650, AD-20652, AD-20648 820CUCAGCACAUUC 2412 GAAUGUGCUGAG 2961 AD-20653, AD-20650, AD-20652 821UCAGCACAUUCC 2413 GGAAUGUGCUGA 2962 AD-20653, AD-20650, AD-20652 822CAGCACAUUCCA 2414 UGGAAUGUGCUG 2963 AD-20653, AD-20652, AD-20658 823AGCACAUUCCAU 2415 AUGGAAUGUGCU 2964 AD-20659, AD-20653, AD-20652,AD-20658 824 GCACAUUCCAUG 2416 CAUGGAAUGUGC 2965 AD-20659, AD-20660,AD-20653, AD-20658 825 CACAUUCCAUGC 2417 GCAUGGAAUGUG 2966 AD-20659,AD-20660, AD-20661, AD-20658 826 ACAUUCCAUGCC 2418 GGCAUGGAAUGU 2967AD-20659, AD-20660, AD-20284, AD-20661, AD-20658 827 CAUUCCAUGCCC 2419GGGCAUGGAAUG 2968 AD-20659, AD-20660, AD-20284, AD-20661, AD-20658 828AUUCCAUGCCCA 2420 UGGGCAUGGAAU 2969 AD-20659, AD-20660, AD-20284,AD-20661, AD-20658 829 UUCCAUGCCCAA 2421 UUGGGCAUGGAA 2970 AD-20659,AD-20660, AD-20284, AD-20661, AD-20658 830 UCCAUGCCCAAG 2422CUUGGGCAUGGA 2971 AD-20659, AD-20660, AD-20284, AD-20661 831CCAUGCCCAAGU 2423 ACUUGGGCAUGG 2972 AD-20660, AD-20284, AD-20661 832CAUGCCCAAGUA 2424 UACUUGGGCAUG 2973 AD-20284, AD-20661 847 CCGGCAGUUCUC2425 GAGAACUGCCGG 2974 AD-20662, AD-20868 848 CGGCAGUUCUCC 2426GGAGAACUGCCG 2975 AD-20663, AD-20868 849 GGCAGUUCUCCC 2427 GGGAGAACUGCC2976 AD-20664, AD-20868 850 GCAGUUCUCCCU 2428 AGGGAGAACUGC 2977AD-20665, AD-20868 851 CAGUUCUCCCUG 2429 CAGGGAGAACUG 2978 AD-20666,AD-20868 852 AGUUCUCCCUGG 2430 CCAGGGAGAACU 2979 AD-20666, AD-20868 853GUUCUCCCUGGA 2431 UCCAGGGAGAAC 2980 AD-20666, AD-20868 854 UUCUCCCUGGAG2432 CUCCAGGGAGAA 2981 AD-20666, AD-20868 855 UCUCCCUGGAGC 2433GCUCCAGGGAGA 2982 AD-20666, AD-20664 856 CUCCCUGGAGCA 2434 UGCUCCAGGGAG2983 AD-20666, AD-20671 857 UCCCUGGAGCAC 2435 GUGCUCCAGGGA 2984AD-20672, AD-20671 858 CCCUGGAGCACG 2436 CGUGCUCCAGGG 2985 AD-20672,AD-20671 859 CCUGGAGCACGU 2437 ACGUGCUCCAGG 2986 AD-20672, AD-20671 860CUGGAGCACGUC 2438 GACGUGCUCCAG 2987 AD-20672, AD-20671 861 UGGAGCACGUCC2439 GGACGUGCUCCA 2988 AD-20672, AD-20671, AD-20676 862 GGAGCACGUCCA2440 UGGACGUGCUCC 2989 AD-20672, AD-20671, AD-20676 863 GAGCACGUCCAC2441 GUGGACGUGCUC 2990 AD-20672, AD-20671, AD-20676 864 AGCACGUCCACG2442 CGUGGACGUGCU 2991 AD-20672, AD-20676 865 GCACGUCCACGG 2443CCGUGGACGUGC 2992 AD-20675, AD-20676 866 CACGUCCACGGC 2444 GCCGUGGACGUG2993 AD-20675, AD-20676 867 ACGUCCACGGCU 2445 AGCCGUGGACGU 2994AD-20675, AD-20676 965 UCCGACAUCACC 2446 GGUGAUGUCGGA 2995 AD-20679,AD-20678 966 CCGACAUCACCG 2447 CGGUGAUGUCGG 2996 AD-20680, AD-20678 967CGACAUCACCGA 2448 UCGGUGAUGUCG 2997 AD-20681, AD-20678 968 GACAUCACCGAG2449 CUCGGUGAUGUC 2998 AD-20678, AD-20682 969 ACAUCACCGAGC 2450GCUCGGUGAUGU 2999 AD-20678, AD-20682 970 CAUCACCGAGCU 2451 AGCUCGGUGAUG3000 AD-20678, AD-20682 971 AUCACCGAGCUG 2452 CAGCUCGGUGAU 3001AD-20678, AD-20682 972 UCACCGAGCUGG 2453 CCAGCUCGGUGA 3002 AD-20686,AD-20682 973 CACCGAGCUGGC 2454 GCCAGCUCGGUG 3003 AD-20686, AD-20687 974ACCGAGCUGGCU 2455 AGCCAGCUCGGU 3004 AD-20686, AD-20687 975 CCGAGCUGGCUC2456 GAGCCAGCUCGG 3005 AD-20686, AD-20689 976 CGAGCUGGCUCC 2457GGAGCCAGCUCG 3006 AD-20686, AD-20689 977 GAGCUGGCUCCU 2458 AGGAGCCAGCUC3007 AD-20686, AD-20689 978 AGCUGGCUCCUG 2459 CAGGAGCCAGCU 3008AD-20686, AD-20689 979 GCUGGCUCCUGC 2460 GCAGGAGCCAGC 3009 AD-20686,AD-20689 980 CUGGCUCCUGCC 2461 GGCAGGAGCCAG 3010 AD-20691, AD-20689 981UGGCUCCUGCCA 2462 UGGCAGGAGCCA 3011 AD-20691, AD-20689 982 GGCUCCUGCCAG2463 CUGGCAGGAGCC 3012 AD-20691, AD-20689 983 GCUCCUGCCAGC 2464GCUGGCAGGAGC 3013 AD-20692, AD-20691 984 CUCCUGCCAGCC 2465 GGCUGGCAGGAG3014 AD-20692, AD-20691 1011 GCGGGAGCAUAG 2466 CUAUGCUCCCGC 3015AD-20693, AD-20694 1012 CGGGAGCAUAGA 2467 UCUAUGCUCCCG 3016 AD-20693,AD-20694 1013 GGGAGCAUAGAC 2468 GUCUAUGCUCCC 3017 AD-20693, AD-206941014 GGAGCAUAGACG 2469 CGUCUAUGCUCC 3018 AD-20693, AD-20694 1015GAGCAUAGACGA 2470 UCGUCUAUGCUC 3019 AD-20693, AD-20694 1016 AGCAUAGACGAG2471 CUCGUCUAUGCU 3020 AD-20693, AD-20694 1017 GCAUAGACGAGA 2472UCUCGUCUAUGC 3021 AD-20693, AD-20694 1018 CAUAGACGAGAG 2473 CUCUCGUCUAUG3022 AD-20698, AD-20694 1019 AUAGACGAGAGG 2474 CCUCUCGUCUAU 3023AD-20697, AD-20698 1020 UAGACGAGAGGC 2475 GCCUCUCGUCUA 3024 AD-20697,AD-20698 1021 AGACGAGAGGCC 2476 GGCCUCUCGUCU 3025 AD-20697, AD-206981048 CCUGGUGCGUGU 2477 ACACGCACCAGG 3026 AD-20700, AD-20699 1049CUGGUGCGUGUC 2478 GACACGCACCAG 3027 AD-20700, AD-20699 1050 UGGUGCGUGUCA2479 UGACACGCACCA 3028 AD-20700, AD-20702 1051 GGUGCGUGUCAA 2480UUGACACGCACC 3029 AD-20700, AD-20702 1052 GUGCGUGUCAAG 2481 CUUGACACGCAC3030 AD-20700, AD-20703, AD-20702 1053 UGCGUGUCAAGG 2482 CCUUGACACGCA3031 AD-20700, AD-20703, AD-20702 1054 GCGUGUCAAGGA 2483 UCCUUGACACGC3032 AD-20700, AD-20703, AD-20702 1055 CGUGUCAAGGAG 2484 CUCCUUGACACG3033 AD-20700, AD-20703, AD-20702 1056 GUGUCAAGGAGG 2485 CCUCCUUGACAC3034 AD-20703, AD-20702 1057 UGUCAAGGAGGA 2486 UCCUCCUUGACA 3035AD-20703, AD-20702 1058 GUCAAGGAGGAG 2487 CUCCUCCUUGAC 3036 AD-20705,AD-20703 1059 UCAAGGAGGAGC 2488 GCUCCUCCUUGA 3037 AD-20705, AD-207031060 CAAGGAGGAGCC 2489 GGCUCCUCCUUG 3038 AD-20705, AD-20704 1329ACUUGGAUGCUA 2490 UAGCAUCCAAGU 3039 AD-20707, AD-20706 1330 CUUGGAUGCUAU2491 AUAGCAUCCAAG 3040 AD-20707, AD-20708 1331 UUGGAUGCUAUG 2492CAUAGCAUCCAA 3041 AD-20707, AD-20709 1332 UGGAUGCUAUGG 2493 CCAUAGCAUCCA3042 AD-20707, AD-20710, AD-20709 1333 GGAUGCUAUGGA 2494 UCCAUAGCAUCC3043 AD-20707, AD-20710, AD-20709 1334 GAUGCUAUGGAC 2495 GUCCAUAGCAUC3044 AD-20707, AD-20710, AD-20709 1335 AUGCUAUGGACU 2496 AGUCCAUAGCAU3045 AD-20713, AD-20707, AD-20710, AD-20709 1336 UGCUAUGGACUC 2497GAGUCCAUAGCA 3046 AD-20713, AD-20714, AD-20707, AD-20710, AD-20709 1337GCUAUGGACUCC 2498 GGAGUCCAUAGC 3047 AD-20713, AD-20714, AD-20710,AD-20709 1338 CUAUGGACUCCA 2499 UGGAGUCCAUAG 3048 AD-20713, AD-20714,AD-20710, AD-20709, AD-20716 1339 UAUGGACUCCAA 2500 UUGGAGUCCAUA 3049AD-20713, AD-20714, AD-20710 1340 AUGGACUCCAAC 2501 GUUGGAGUCCAU 3050AD-20713, AD-20714 1341 UGGACUCCAACC 2502 GGUUGGAGUCCA 3051 AD-20713,AD-20714 1342 GGACUCCAACCU 2503 AGGUUGGAGUCC 3052 AD-20713, AD-207141343 GACUCCAACCUG 2504 CAGGUUGGAGUC 3053 AD-20714, AD-20716 1344ACUCCAACCUGG 2505 CCAGGUUGGAGU 3054 AD-20716, AD-20715 1359 ACCUGCAGACCA2506 UGGUCUGCAGGU 3055 AD-20717, AD-20718 1360 CCUGCAGACCAU 2507AUGGUCUGCAGG 3056 AD-20717, AD-20718 1361 CUGCAGACCAUG 2508 CAUGGUCUGCAG3057 AD-20720, AD-20718 1362 UGCAGACCAUGC 2509 GCAUGGUCUGCA 3058AD-20718, AD-20721 1363 GCAGACCAUGCU 2510 AGCAUGGUCUGC 3059 AD-20718,AD-20722 1364 CAGACCAUGCUG 2511 CAGCAUGGUCUG 3060 AD-20718, AD-207221365 AGACCAUGCUGA 2512 UCAGCAUGGUCU 3061 AD-20718, AD-20722 1366GACCAUGCUGAG 2513 CUCAGCAUGGUC 3062 AD-20718, AD-20722 1367 ACCAUGCUGAGC2514 GCUCAGCAUGGU 3063 AD-20726, AD-20722 1368 CCAUGCUGAGCA 2515UGCUCAGCAUGG 3064 AD-20727, AD-20722 1369 CAUGCUGAGCAG 2516 CUGCUCAGCAUG3065 AD-20728, AD-20722 1370 AUGCUGAGCAGC 2517 GCUGCUCAGCAU 3066AD-20728, AD-20722 1371 UGCUGAGCAGCC 2518 GGCUGCUCAGCA 3067 AD-20728,AD-20730 1372 GCUGAGCAGCCA 2519 UGGCUGCUCAGC 3068 AD-20728, AD-207301373 CUGAGCAGCCAC 2520 GUGGCUGCUCAG 3069 AD-20728, AD-20730 1374UGAGCAGCCACG 2521 CGUGGCUGCUCA 3070 AD-20728, AD-20730 1375 GAGCAGCCACGG2522 CCGUGGCUGCUC 3071 AD-20728, AD-20730 1376 AGCAGCCACGGC 2523GCCGUGGCUGCU 3072 AD-20728, AD-20730 1377 GCAGCCACGGCU 2524 AGCCGUGGCUGC3073 AD-20733, AD-20730 1378 CAGCCACGGCUU 2525 AAGCCGUGGCUG 3074AD-20733, AD-20730 1379 AGCCACGGCUUC 2526 GAAGCCGUGGCU 3075 AD-20733,AD-20735 1380 GCCACGGCUUCA 2527 UGAAGCCGUGGC 3076 AD-20733, AD-207361381 CCACGGCUUCAG 2528 CUGAAGCCGUGG 3077 AD-20733, AD-20736 1382CACGGCUUCAGC 2529 GCUGAAGCCGUG 3078 AD-20738, AD-20736 1383 ACGGCUUCAGCG2530 CGCUGAAGCCGU 3079 AD-20739, AD-20736 1384 CGGCUUCAGCGU 2531ACGCUGAAGCCG 3080 AD-20739, AD-20740 1385 GGCUUCAGCGUG 2532 CACGCUGAAGCC3081 AD-20741, AD-20740 1386 GCUUCAGCGUGG 2533 CCACGCUGAAGC 3082AD-20742, AD-20741 1387 CUUCAGCGUGGA 2534 UCCACGCUGAAG 3083 AD-20742,AD-20741 1388 UUCAGCGUGGAC 2535 GUCCACGCUGAA 3084 AD-20742, AD-207411389 UCAGCGUGGACA 2536 UGUCCACGCUGA 3085 AD-20742, AD-20741 1390CAGCGUGGACAC 2537 GUGUCCACGCUG 3086 AD-20742, AD-20741 1391 AGCGUGGACACC2538 GGUGUCCACGCU 3087 AD-20742, AD-20741 1392 GCGUGGACACCA 2539UGGUGUCCACGC 3088 AD-20742, AD-20741 1407 CCCUGCUGGACC 2540 GGUCCAGCAGGG3089 AD-20744, AD-20743 1408 CCUGCUGGACCU 2541 AGGUCCAGCAGG 3090AD-20744, AD-20743 1409 CUGCUGGACCUG 2542 CAGGUCCAGCAG 3091 AD-20746,AD-20743 1410 UGCUGGACCUGU 2543 ACAGGUCCAGCA 3092 AD-20746, AD-207431411 GCUGGACCUGUU 2544 AACAGGUCCAGC 3093 AD-20746, AD-20743 1412CUGGACCUGUUC 2545 GAACAGGUCCAG 3094 AD-20746, AD-20743 1413 UGGACCUGUUCA2546 UGAACAGGUCCA 3095 AD-20746, AD-20743 1414 GGACCUGUUCAG 2547CUGAACAGGUCC 3096 AD-20746, AD-20747 1415 GACCUGUUCAGC 2548 GCUGAACAGGUC3097 AD-20746, AD-20747 1416 ACCUGUUCAGCC 2549 GGCUGAACAGGU 3098AD-20746, AD-20747 1428 CCUCGGUGACCG 2550 CGGUCACCGAGG 3099 AD-20749,AD-20748 1429 CUCGGUGACCGU 2551 ACGGUCACCGAG 3100 AD-20749, AD-207501430 UCGGUGACCGUG 2552 CACGGUCACCGA 3101 AD-20751, AD-20750 1431CGGUGACCGUGC 2553 GCACGGUCACCG 3102 AD-20751, AD-20752 1432 GGUGACCGUGCC2554 GGCACGGUCACC 3103 AD-20751, AD-20753 1433 GUGACCGUGCCC 2555GGGCACGGUCAC 3104 AD-20751, AD-20754 1434 UGACCGUGCCCG 2556 CGGGCACGGUCA3105 AD-20751, AD-20755 1435 GACCGUGCCCGA 2557 UCGGGCACGGUC 3106AD-20751, AD-20756 1436 ACCGUGCCCGAC 2558 GUCGGGCACGGU 3107 AD-20751,AD-20757 1437 CCGUGCCCGACA 2559 UGUCGGGCACGG 3108 AD-20751, AD-207581438 CGUGCCCGACAU 2560 AUGUCGGGCACG 3109 AD-20759, AD-20758 1439GUGCCCGACAUG 2561 CAUGUCGGGCAC 3110 AD-20760, AD-20759 1440 UGCCCGACAUGA2562 UCAUGUCGGGCA 3111 AD-20760, AD-20761 1441 GCCCGACAUGAG 2563CUCAUGUCGGGC 3112 AD-20760, AD-20762 1442 CCCGACAUGAGC 2564 GCUCAUGUCGGG3113 AD-20763, AD-20762 1443 CCGACAUGAGCC 2565 GGCUCAUGUCGG 3114AD-20763, AD-20764 1444 CGACAUGAGCCU 2566 AGGCUCAUGUCG 3115 AD-20763,AD-20764 1445 GACAUGAGCCUG 2567 CAGGCUCAUGUC 3116 AD-20763, AD-207641446 ACAUGAGCCUGC 2568 GCAGGCUCAUGU 3117 AD-20763, AD-20764 1447CAUGAGCCUGCC 2569 GGCAGGCUCAUG 3118 AD-20768, AD-20764 1448 AUGAGCCUGCCU2570 AGGCAGGCUCAU 3119 AD-20768, AD-20764 1449 UGAGCCUGCCUG 2571CAGGCAGGCUCA 3120 AD-20768, AD-20764 1450 GAGCCUGCCUGA 2572 UCAGGCAGGCUC3121 AD-20771, AD-20764 1451 AGCCUGCCUGAC 2573 GUCAGGCAGGCU 3122AD-20771, AD-20772 1452 GCCUGCCUGACC 2574 GGUCAGGCAGGC 3123 AD-20771,AD-20773 1453 CCUGCCUGACCU 2575 AGGUCAGGCAGG 3124 AD-20771, AD-207741454 CUGCCUGACCUU 2576 AAGGUCAGGCAG 3125 AD-20771, AD-20775 1455UGCCUGACCUUG 2577 CAAGGUCAGGCA 3126 AD-20771, AD-20776 1456 GCCUGACCUUGA2578 UCAAGGUCAGGC 3127 AD-20771, AD-20777 1457 CCUGACCUUGAC 2579GUCAAGGUCAGG 3128 AD-20771, AD-20778 1458 CUGACCUUGACA 2580 UGUCAAGGUCAG3129 AD-20779, AD-20778 1459 UGACCUUGACAG 2581 CUGUCAAGGUCA 3130AD-20779, AD-20780 1460 GACCUUGACAGC 2582 GCUGUCAAGGUC 3131 AD-20779,AD-20780 1461 ACCUUGACAGCA 2583 UGCUGUCAAGGU 3132 AD-20779, AD-207801462 CCUUGACAGCAG 2584 CUGCUGUCAAGG 3133 AD-20779, AD-20780 1463CUUGACAGCAGC 2585 GCUGCUGUCAAG 3134 AD-20779, AD-20780 1464 UUGACAGCAGCC2586 GGCUGCUGUCAA 3135 AD-20779, AD-20780 1465 UGACAGCAGCCU 2587AGGCUGCUGUCA 3136 AD-20779, AD-20780 1466 GACAGCAGCCUG 2588 CAGGCUGCUGUC3137 AD-20781, AD-20780 1467 ACAGCAGCCUGG 2589 CCAGGCUGCUGU 3138AD-20781, AD-20782 1482 GUAUCCAAGAGC 2590 GCUCUUGGAUAC 3139 AD-20783,AD-20784 1483 UAUCCAAGAGCU 2591 AGCUCUUGGAUA 3140 AD-20783, AD-20785,AD-20784 1484 AUCCAAGAGCUC 2592 GAGCUCUUGGAU 3141 AD-20783, AD-20785,AD-20784 1485 UCCAAGAGCUCC 2593 GGAGCUCUUGGA 3142 AD-20783, AD-20785,AD-20784 1486 CCAAGAGCUCCU 2594 AGGAGCUCUUGG 3143 AD-20783, AD-20785,AD-20784 1487 CAAGAGCUCCUG 2595 CAGGAGCUCUUG 3144 AD-20783, AD-20785,AD-20784 1488 AAGAGCUCCUGU 2596 ACAGGAGCUCUU 3145 AD-20783, AD-20785,AD-20784 1489 AGAGCUCCUGUC 2597 GACAGGAGCUCU 3146 AD-20785, AD-207841490 GAGCUCCUGUCU 2598 AGACAGGAGCUC 3147 AD-20786, AD-20785 1491AGCUCCUGUCUC 2599 GAGACAGGAGCU 3148 AD-20786, AD-20788 1492 GCUCCUGUCUCC2600 GGAGACAGGAGC 3149 AD-20788, AD-20787 1547 GAUUCAGGGAAG 2601CUUCCCUGAAUC 3150 AD-20789, AD-20790 1548 AUUCAGGGAAGC 2602 GCUUCCCUGAAU3151 AD-20789, AD-20791 1549 UUCAGGGAAGCA 2603 UGCUUCCCUGAA 3152AD-20789, AD-20791 1550 UCAGGGAAGCAG 2604 CUGCUUCCCUGA 3153 AD-20789,AD-20791 1551 CAGGGAAGCAGC 2605 GCUGCUUCCCUG 3154 AD-20789, AD-207911552 AGGGAAGCAGCU 2606 AGCUGCUUCCCU 3155 AD-20789, AD-20791 1553GGGAAGCAGCUG 2607 CAGCUGCUUCCC 3156 AD-20789, AD-20791 1554 GGAAGCAGCUGG2608 CCAGCUGCUUCC 3157 AD-20790, AD-20791 1602 CCGGCUCCGUGG 2609CCACGGAGCCGG 3158 AD-20793, AD-20792 1603 CGGCUCCGUGGA 2610 UCCACGGAGCCG3159 AD-20793, AD-20792 1604 GGCUCCGUGGAC 2611 GUCCACGGAGCC 3160AD-20795, AD-20792 1605 GCUCCGUGGACA 2612 UGUCCACGGAGC 3161 AD-20795,AD-20792 1606 CUCCGUGGACAC 2613 GUGUCCACGGAG 3162 AD-20795, AD-208701607 UCCGUGGACACC 2614 GGUGUCCACGGA 3163 AD-20871, AD-20870 1608CCGUGGACACCG 2615 CGGUGUCCACGG 3164 AD-20871, AD-20870 1609 CGUGGACACCGG2616 CCGGUGUCCACG 3165 AD-20871, AD-20870 1610 GUGGACACCGGG 2617CCCGGUGUCCAC 3166 AD-20871, AD-20870 1611 UGGACACCGGGA 2618 UCCCGGUGUCCA3167 AD-20871, AD-20870 1612 GGACACCGGGAG 2619 CUCCCGGUGUCC 3168AD-20871, AD-20870 1613 GACACCGGGAGC 2620 GCUCCCGGUGUC 3169 AD-20871,AD-20870 1633 GCCGGUGCUGUU 2621 AACAGCACCGGC 3170 AD-20797, AD-208721634 CCGGUGCUGUUU 2622 AAACAGCACCGG 3171 AD-20872, AD-20798 1635CGGUGCUGUUUG 2623 CAAACAGCACCG 3172 AD-20872, AD-20799 1636 GGUGCUGUUUGA2624 UCAAACAGCACC 3173 AD-20872, AD-20799 1637 GUGCUGUUUGAG 2625CUCAAACAGCAC 3174 AD-20872, AD-20799 1638 UGCUGUUUGAGC 2626 GCUCAAACAGCA3175 AD-20872, AD-20799 1639 GCUGUUUGAGCU 2627 AGCUCAAACAGC 3176AD-20872, AD-20799 1640 CUGUUUGAGCUG 2628 CAGCUCAAACAG 3177 AD-20798,AD-20799 1641 UGUUUGAGCUGG 2629 CCAGCUCAAACA 3178 AD-20798, AD-207991698 CCACCAUCUCCC 2630 GGGAGAUGGUGG 3179 AD-20873, AD-20800 1699CACCAUCUCCCU 2631 AGGGAGAUGGUG 3180 AD-20801, AD-20800 1700 ACCAUCUCCCUG2632 CAGGGAGAUGGU 3181 AD-20801, AD-20800 1701 CCAUCUCCCUGC 2633GCAGGGAGAUGG 3182 AD-20801, AD-20800 1702 CAUCUCCCUGCU 2634 AGCAGGGAGAUG3183 AD-20801, AD-20800 1703 AUCUCCCUGCUG 2635 CAGCAGGGAGAU 3184AD-20801, AD-20800 1704 UCUCCCUGCUGA 2636 UCAGCAGGGAGA 3185 AD-20801,AD-20800 1705 CUCCCUGCUGAC 2637 GUCAGCAGGGAG 3186 AD-20801, AD-208002009 CAGGUUGUUCAU 2638 AUGAACAACCUG 3187 AD-20279, AD-20278 2010AGGUUGUUCAUA 2639 UAUGAACAACCU 3188 AD-20279, AD-20280 2011 GGUUGUUCAUAG2640 CUAUGAACAACC 3189 AD-20279, AD-20278 2012 GUUGUUCAUAGU 2641ACUAUGAACAAC 3190 AD-20279, AD-20280 2013 UUGUUCAUAGUC 2642 GACUAUGAACAA3191 AD-20279, AD-20278 2014 UGUUCAUAGUCA 2643 UGACUAUGAACA 3192AD-20279, AD-20280 2015 GUUCAUAGUCAG 2644 CUGACUAUGAAC 3193 AD-20279,AD-20278 2016 UUCAUAGUCAGA 2645 UCUGACUAUGAA 3194 AD-20279, AD-20280

Example 2 Preparation of siRNAs

Small scale synthesis is used to prepare HSF1 siRNAs; medium and largescale syntheses can also be used to prepare these siRNAs in largerquantities.

Small Scale Synthesis and Purification Methods for the Initial Screens(1 μmole Scale).

Small scale synthesis is used to generate siRNAs.

HSF1 sequences are synthesized on MerMade 192 synthesizer(BioAutomation, Plano, Tex.) at 1 μmol scale.

For all the sequences in the list, endolight' chemistry is applied asdetailed below:

All pyrimidines (cytosine and uridine) in the sense strand contain2′-O-Methyl bases (2′ O-Methyl C and 2′-O-Methyl U).

In the antisense strand, pyrimidines adjacent to (towards 5′ position)ribo A nucleoside are replaced with their corresponding 2-O-Methylnucleosides.

A two base dTdT extension at 3′ end of both sense and antisensesequences is introduced.

The sequence file is converted to a text file to make it compatible forloading in the MerMade 192 synthesis software.

Synthesis, Cleavage and Deprotection:

The synthesis of HSF1 sequences can use solid supported oligonucleotidesynthesis using phosphoramidite chemistry.

The synthesis of the above sequences is performed at 1 um scale in 96well plates. The ribo and 2-O-Methyl phosphoramidite solutions areprepared at 0.1M concentration and ethyl thio tetrazole (0.6M inAcetonitrile) is used as activator. Deblock solution, oxidizer solutionand capping solution are prepared according to standard processes.

The synthesized sequences are cleaved and deprotected in 96 well plates,using methylamine solution (a 3:1 mixture of aqueous and ethanolicsolutions) in the first step and fluoride reagent in the second step.The crude sequences are precipitated using acetone: ethanol (80:20) mixand the pellet are re-suspended in 0.02M sodium acetate buffer. Samplesfrom each sequence are analyzed by LC-MS to confirm the identity, UV forquantification and a selected set of samples by IEX chromatography todetermine purity.

Purification and Desalting:

HSF1 tiled sequences are purified on AKTA explorer purification systemusing Source 15Q column. A column temperature of 65C is maintainedduring purification. Sample injection and collection are performed in 96well (1.8mL -deep well) plates. A single peak corresponding to the fulllength sequence is collected in the eluent. The purified sequences aredesalted on a Sephadex G25 column using AKTA purifier. The concentrationof desalted HSF1 sequences are calculated using absorbance at 260 nmwavelength and purity was measured by ion exchange chromatography.

Annealing:

Purified desalted sense and antisense single strands are mixed inequimolar amounts and annealed to form HSF1 duplexes. The duplexes areprepared at 10 uM concentration in 1× PBS buffer and tested by capillarygel electrophoresis for purity.

Medium Scale Synthesis and Purification (1-50 μmol)

Medium scale synthesis can also be used to generate siRNAs.

Single-stranded RNAs in scales between 1 and 50 μmol are prepared bysolid phase synthesis using an ABI DNA/RNA Synthesizer 394 (AppliedBiosystems) and controlled pore glass (CPG, 500 Å, loading 80-100μmol/g) purchased from Prime Synthesis (Aston, Pa.) as the solidsupport. For larger scales, empty synthesis columns (10 μmol) from GlenResearch Corp. and large amidite (80 mL) and reagent bottles (450 mL)are used. RNA and RNA containing 2μ-O-methyl nucleotides are generatedby solid phase synthesis employing the corresponding phosphoramiditesand 2 μ-O-methyl phosphoramidites, respectively (ChemGenes, Wilmington,Mass.). These building blocks are incorporated at selected sites withinthe sequence of the oligoribonucleotide chain using standard nucleosidephosphoramidite chemistry such as described in Current Protocols inNucleic Acid Chemistry, Beaucage, S. L. et al. (Edrs.), John Wiley &Sons, Inc., New York, N.Y., USA. Phosphorothioate linkages areintroduced using a solution of the 0.1 M DDTT (AM Chemicals, Oceanside,Calif.) in pyridine. Further ancillary reagents are obtained from GlenResearch Corp. (Sterling, Va.).

Deprotection and purification of the crude oligoribonucleotides by anionexchange HPLC are carried out according to established procedures.Yields and concentrations are determined spectrophotometrically at awavelength of 260 nm. Double stranded RNA is generated by mixing anequimolar solution of complementary strands in annealing buffer(typically phosphate buffered solution, PBS, Ambion, Applied Biosystems,Austin, Tex.) at the desired concentration. The mixture is then heatedin a water bath at 85-90° C. for 5 minutes and cooled to roomtemperature over a period of 3-4 hours. The RNA duplex is stored at −20°C. until use.

Example 3 HSF1 RNAi Agents

HSF1 siRNAs are provided in Table 2, and prepared as described inExample 2. The activity of these RNAi agents is listed in Table 4.

Table 4 indicates the residual level of HSF1 expression in approximately20,000 W138 or HeLa cells treated with 10 nM of the indicated siRNA.Expression measurements are done 24 hours after transfection. Expressionis measured using RT-qRT [real-time quantitative reverse transcription].The residual activity is normalized to HSF1 expression in Luc siRNAtransfected cells, and 1.000000=100% gene expression relative to thecontrol, or no or 0% gene knockdown; and 0.000000=0% gene expression, orcomplete or 100% gene knockdown in W138 or HeLa cells. A low number(closer to zero) indicates a more potent siRNA. For example, the “WI38”column for siRNA AD-20303 indicates “0.056232” meaning that the residualgene activity is 5.6%, or 94.4% gene knock-down at 10 nM. In the HeLacolumn, “0.098871432” indicates approximately 9.9% residual activity, or90.1% gene knockdown at 10 nM.

A serial dilution of the siRNAs was performed and the data curve fit tocalculate the dose (concentration) required to knock down geneexpression by 50% (“EC50,” or effective concentration estimated toreduce gene expression by 50%). The “EC50 Average” indicates the amountof siRNA in nM in which 50% gene knockdown is expected to be achieved;the given number is an average of two experiments. A lower numberindicates a more potent siRNA. A blank cell indicates that no siRNA withthat sequence was tested in that particular test, or that such data isnot included herein.

In Table 4, the suffixes “-b1”, “-b2”, etc., indicate batch numbers.Thus “b1” is from batch 1, “b2” is from batch 2, etc. Thus, for example,“AD-20489-b1”, “AD-20489-b2” and “AD-20489” all have the same sequenceand are chemically identical.

TABLE 4 ACTIVITY OF HSF1 RNAi AGENTS Sense 5′-3′ Antisense 5′-3′ EC50Duplex modified modified average Name Position SEQ ID NO SEQ ID NO WI38(10 nM) HELA (10 nM) (nM)* AD-20284 827 1350 1862 0.543633 1.057018041AD-20285 212 1036 1548 0.914447 1.641966293 AD-20286 213 1037 15490.813034 1.375673997 AD-20287 216 1038 1550 0.696869 1.375574862AD-20288 217 1039 1551 0.662908 1.198888545 AD-20289 218 1040 15520.862919 1.474587971 AD-20290 219 1041 1553 0.853455 1.202220643AD-20291 220 1042 1554 0.747856 1.168784268 AD-20292 221 1043 15550.898657 0.972865247 AD-20293 222 1044 1556 0.824067 1.228130423AD-20294 223 1045 1557 0.521985 0.592834658 AD-20295 224 1046 15580.987656 1.414757168 AD-20296 225 1047 1559 0.195372 0.369849098AD-20297 226 1048 1560 0.635752 0.861551334 AD-20298 227 1049 15610.615987 1.094714334 AD-20299 228 1050 1562 0.730146 1.266851398AD-20300 229 1051 1563 0.385889 0.479920139 AD-20301 230 1052 15640.553587 0.629050308 AD-20302 231 1053 1565 0.763571 1.310519322AD-20303 232 1054 1566 0.056232 0.098871432 0.03675 AD-20304 233 10551567 0.631087 1.049741901 AD-20305 234 1056 1568 0.963295 1.205807828AD-20306 235 1057 1569 0.727406 0.806719271 AD-20307 236 1058 15700.959911 0.796014304 AD-20308 237 1059 1571 0.895564 1.676467495AD-20309 238 1060 1572 0.921272 1.144712192 AD-20310 239 1061 15730.88654 1.038915255 AD-20311 240 1062 1574 0.722451 0.963062743 AD-20312241 1063 1575 0.256473 0.406750757 AD-20313 242 1064 1576 0.0904040.127185725 0.1542 AD-20314 243 1065 1577 0.707532 0.912250883 AD-20315244 1066 1578 0.188305 0.220760574 0.00639 AD-20316 245 1067 15790.413665 0.74538006 AD-20317 246 1068 1580 0.860383 1.079643078 AD-20318247 1069 1581 0.740659 1.017485803 AD-20319 270 1073 1585 0.7252421.109809374 AD-20320 271 1074 1586 0.656347 0.829339468 AD-20344- 3061075 1587 0.370436 0.484671104 b1 AD-20345- 307 1076 1588 0.3378130.314374139 b1 AD-20346- 309 1078 1590 0.908065 1.342645071 b1 AD-20347-310 1079 1591 0.785716 1.315013835 b1 AD-20348- 311 1080 1592 0.1106170.173142843 0.78766 b1 AD-20349- 312 1081 1593 0.330099 0.600908235 b1AD-20350- 313 1082 1594 0.601169 0.966029148 b1 AD-20351- 314 1083 15950.859247 1.015933117 b1 AD-20352- 315 1084 1596 0.755786 0.724475428 b1AD-20353- 316 1085 1597 0.321073 0.349284689 b1 AD-20354- 317 1086 15980.441617 0.790060291 b1 AD-20355- 318 1087 1599 0.504071 1.043091893 b1AD-20356- 319 1088 1600 0.487357 0.867585579 b1 AD-20357- 320 1089 16010.865058 1.366171522 b1 AD-20358- 321 1090 1602 1.033327 1.291294086 b1AD-20359- 322 1091 1603 1.021823 1.235485413 b1 AD-20360- 323 1092 16040.890403 1.404984726 b1 AD-20361- 324 1093 1605 0.946695 1.257013375 b1AD-20362- 325 1094 1606 0.201283 0.252614379 0.8636 b1 AD-20363- 3261095 1607 0.727406 1.284936927 b1 AD-20364- 327 1096 1608 0.164270.23101335 0.07445 b1 AD-20365- 328 1097 1609 0.155749 0.1280888320.0134 b1 AD-20366- 329 1098 1610 0.20747 0.172548995 0.26565 b1AD-20367- 330 1099 1611 0.633259 0.801146854 b1 AD-20368- 331 1100 16120.622818 1.054126591 b1 AD-20369- 351 1101 1613 0.753131 1.069508982 b1AD-20370- 352 1102 1614 0.774695 0.952729539 b1 AD-20371- 353 1103 16150.48181 0.942790198 b1 AD-20372- 354 1104 1616 0.674589 1.028113827 b1AD-20373- 355 1105 1617 0.097819 0.104764119 0.02475 b1 AD-20374- 3561106 1618 0.347849 0.543484914 b1 AD-20375- 357 1107 1619 0.6383990.66208024 b1 AD-20376- 358 1108 1620 0.050815 0.067088855 0.00507 b1AD-20378- 360 1109 1621 1.035963 0.966029148 0.61274 b1 AD-20379- 3611110 1622 0.454065 0.572640533 b1 AD-20380- 362 1111 1623 0.252760.543419645 b1 AD-20381- 363 1112 1624 1.149473 1.660591875 b1 AD-20382-364 1113 1625 0.950581 1.296847343 b1 AD-20383- 365 1114 1626 0.8477431.278993262 b1 AD-20384- 366 1115 1627 0.862049 1.144731035 b1 AD-20385-367 1116 1628 0.605641 0.829638325 b1 AD-20386- 436 1117 1629 0.0302030.061237298 0.00049 b1 AD-20387- 489 1139 1651 0.285437 0.325718495 b1AD-20388- 490 1140 1652 0.391351 0.505908575 b1 AD-20389- 491 1141 16530.067492 0.082477169 0.07576 b1 AD-20390- 492 1142 1654 0.296460.485136885 b1 AD-20391- 493 1143 1655 0.032161 0.06448988 0.00463 b1AD-20392- 494 1144 1656 0.031801 0.093150137 0.04397 b1 AD-20393- 4951145 1657 0.234886 0.390040961 b1 AD-20394- 496 1146 1658 0.9042971.574701063 b1 AD-20395- 497 1147 1659 0.17985 0.44820074 b1 AD-20396-498 1148 1660 0.322909 0.648454825 b1 AD-20397- 499 1149 1661 0.1099570.16678346 0.2323 b1 AD-20398- 500 1150 1662 0.139138 0.229318478 0.4807b1 AD-20399- 501 1151 1663 0.067419 0.070581041 0.01757 b1 AD-20400- 5021152 1664 0.487807 0.67402138 b1 AD-20401- 503 1153 1665 0.0720050.078942641 0.08459 b1 AD-20402- 504 1154 1666 0.04683 0.0672220330.00705 b1 AD-20403- 505 1155 1667 0.033648 0.040950778 0.00178 b1AD-20404- 506 1156 1668 0.059783 0.094532132 0.00881 b1 AD-20405- 5091159 1671 1.097465 1.526589204 b1 AD-20406- 510 1160 1672 0.1047310.262088429 0.00878 b1 AD-20407- 511 1161 1673 0.046705 0.1228592150.00183 b1 AD-20408- 512 1162 1674 0.042965 0.099681042 0.00315 b1AD-20409- 513 1163 1675 0.094698 0.183015108 0.00307 b1 AD-20410- 5141164 1676 0.073099 0.092506702 0.015 b1 AD-20411- 515 1165 1677 0.0813830.138821902 0.00193 b1 AD-20412- 516 1166 1678 0.801088 1.025055268 b1AD-20413- 517 2046 2047 0.1237 0.1888102 b1 AD-20414- 518 1167 16790.888671 0.939545319 b1 AD-20415- 519 1168 1680 0.776025 1.010748814 b1AD-20416- 520 1169 1681 0.901076 1.64781519 b1 AD-20417- 521 1170 16820.906995 1.439357047 b1 AD-20418- 522 1171 1683 0.650295 1.400603047 b1AD-20419- 523 1172 1684 0.922855 1.598754699 b1 AD-20420- 524 1173 16850.91692 1.531567195 b1 AD-20421- 525 1174 1686 0.31653 0.557463247 b1AD-20422- 526 1175 1687 0.059576 0.093484193 0.00157 b1 AD-20423- 5271176 1688 0.657285 0.921602755 b1 AD-20424- 528 1177 1689 0.7116140.570505181 b1 AD-20425- 529 1178 1690 1.020643 0.990138123 b1 AD-20426-530 1179 1691 0.200208 0.311228942 b1 AD-20427- 531 1180 1692 0.3645990.453759578 b1 AD-20428- 532 1181 1693 0.131633 0.289178993 1.4762 b1AD-20429- 533 1182 1694 0.481253 0.633425699 b1 AD-20430- 534 1183 16950.8211 1.641493076 b1 AD-20431- 535 1184 1696 0.404548 1.049716684 b1AD-20432- 536 1185 1697 0.494569 0.752912672 b1 AD-20433- 537 1186 16980.308767 0.465128645 b1 AD-20434- 538 1187 1699 0.076144 0.1294858410.00334 b1 AD-20435- 539 1188 1700 0.039875 0.072404187 0.00036 b1AD-20436- 540 1189 1701 0.167895 0.377674844 b1 AD-20437- 541 1190 17020.031574 0.064053646 b1 AD-20438- 542 1191 1703 0.026509 0.0597555530.00007 b1 AD-20439- 543 1192 1704 0.085946 0.113834413 0.0002 b1AD-20487- 544 1193 1705 0.024337 0.060353389 b1 AD-20488- 545 1194 17060.035714 0.070358389 0.00016 b1 AD-20489- 546 1195 1707 0.0303310.057370933 <1E−05 b1 AD-20490- 547 1196 1708 0.0329 0.08714602 0.00021b1 AD-20491- 548 1197 1709 0.034581 0.076846617 0.00004 b1 AD-20492- 5491198 1710 0.407597 0.338288586 b1 AD-20493- 550 1199 1711 0.0365670.094916679 0.00196 b1 AD-20494- 579 1200 1712 0.251648 0.336937859 b1AD-20495- 580 1201 1713 0.073028 0.095448495 0.0207 b1 AD-20496- 5811202 1714 0.619868 0.676154345 b1 AD-20497- 582 1203 1715 0.8700690.843820864 b1 AD-20498- 583 1204 1716 0.711631 1.244198436 b1 AD-20499-584 1205 1717 0.864557 1.565852803 b1 AD-20500- 585 1206 1718 0.5906661.156715971 b1 AD-20501- 586 1207 1719 0.321833 0.63913362 b1 AD-20502-587 1208 1720 0.085336 0.255995345 0.01974 b1 AD-20503- 588 1209 17210.699393 1.425281914 b1 AD-20504- 589 1210 1722 0.375606 0.858565436 b1AD-20505- 590 1211 1723 0.811959 1.357125789 b1 AD-20506- 591 1212 17240.372207 0.80389444 b1 AD-20507- 592 1213 1725 0.032835 0.0695892690.22784 b1 AD-20508- 593 1214 1726 0.617356 1.083293756 b1 AD-20509- 5941215 1727 1.067763 1.559741673 b1 AD-20510- 595 1216 1728 0.2299140.368671667 b1 AD-20511- 596 1217 1729 0.254225 0.637831523 b1 AD-20512-597 1218 1730 0.410688 0.672742156 b1 AD-20513- 598 1219 1731 0.0743270.152299691 0.09514 b1 AD-20514- 660 1220 1732 0.814998 1.553693251 b1AD-20515- 661 1221 1733 0.754914 1.032432979 b1 AD-20516- 662 1222 17340.757584 1.275663046 b1 AD-20517- 663 1223 1735 0.907344 0.937222684 b1AD-20518- 664 1224 1736 0.606761 0.859343664 b1 AD-20519- 665 1225 17370.958802 1.352308689 b1 AD-20520- 666 1226 1738 1.027619 1.548903545 b1AD-20521- 667 1227 1739 1.041889 1.298156014 b1 AD-20522- 668 1228 17401.263989 1.229374539 b1 AD-20523- 669 1229 1741 0.975755 1.708883973 b1AD-20524- 670 1230 1742 0.910412 1.363879889 b1 AD-20525- 671 1231 17430.992543 1.746803026 b1 AD-20526- 672 1232 1744 0.956605 0.951954008 b1AD-20527- 673 1233 1745 0.188065 0.228063688 0.06117 b1 AD-20528- 6741234 1746 0.817921 0.925922775 b1 AD-20529- 675 1235 1747 0.9264091.446572494 b1 AD-20530- 676 1236 1748 0.373532 0.18718473 b1 AD-20531-677 1237 1749 0.471349 0.223370205 b1 AD-20532- 678 1238 1750 1.4048481.377927517 b1 AD-20533- 679 1239 1751 1.285181 1.422003002 b1 AD-20534-680 1240 1752 0.634896 0.56546462 b1 AD-20535- 681 1241 1753 0.1746610.148940901 0.29242 b1 AD-20536- 682 1242 1754 0.803392 0.89969547 b1AD-20537- 683 1243 1755 0.855147 1.158722144 b1 AD-20538- 684 1244 17560.444431 0.500867185 b1 AD-20539- 685 1245 1757 0.879273 1.162891581 b1AD-20540- 690 1250 1762 0.989234 1.313170683 b1 AD-20541- 691 1251 17630.897878 1.030119632 b1 AD-20542- 692 1252 1764 0.481253 0.434545147 b1AD-20543- 693 1253 1765 0.864557 0.511391469 b1 AD-20544- 694 1254 17660.174857 0.052921675 0.00594 b1 AD-20545- 695 1255 1767 0.0715770.047121676 0.00711 b1 AD-20546- 696 1256 1768 0.16775 0.1474755950.2047 b1 AD-20547- 697 1257 1769 0.192017 0.135189447 0.03551 b1AD-20548- 698 1258 1770 0.181681 0.217307804 0.00061 b2 AD-20549- 6991259 1771 0.198941 0.143891617 0.1391 b1 AD-20550- 700 1260 17720.566237 0.542455982 b1 AD-20551- 701 1261 1773 0.82361 1.446572494 b1AD-20552- 702 1262 1774 0.138619 0.128764954 0.08771 b1 AD-20553- 7031263 1775 0.734694 0.767332896 b1 AD-20554- 704 1264 1776 0.5963270.451450159 b1 AD-20555- 705 1265 1777 0.121969 0.126599989 0.02063 b1AD-20556- 706 1266 1778 0.159416 0.071195236 0.00754 b1 AD-20557- 7071267 1779 0.077337 0.04632467 0.00082 b1 AD-20558- 731 1291 18030.156539 0.093590117 0.07236 b1 AD-20559- 732 1292 1804 0.303490.280860279 b1 AD-20560- 733 1293 1805 0.323181 0.206972165 <1E−05 b2AD-20561- 734 1294 1806 0.099423 0.07673151 0.02383 b1 AD-20562- 7351295 1807 0.039811 0.042199399 0.00048 b1 AD-20563- 736 1296 18080.123696 0.188810207 0.00004 b2 AD-20564- 737 1297 1809 0.4489390.206972165 <1E−05 b2 AD-20565- 738 1298 1810 0.060827 0.0552646210.01163 b1 AD-20566- 739 1299 1811 0.228259 0.203420172 0.00268 b2AD-20567- 740 1300 1812 0.852163 0.397272737 b1 AD-20568- 741 1301 18130.975474 1.06284006 b1 AD-20569- 742 1302 1814 1.038248 1.70257293 b1AD-20571- 744 1303 1815 0.914379 0.887564894 b1 AD-20572- 745 1304 18160.07702 0.047860345 0.06155 b1 AD-20573- 746 1305 1817 0.6307830.659280255 b1 AD-20574- 747 2048 2049 0.22826 0.2034202 1.37204 b1AD-20575- 748 1306 1818 0.124712 0.101746738 0.13506 b1 AD-20576- 7491307 1819 0.329885 0.232889354 b1 AD-20577- 750 1308 1820 0.2236270.133103976 0.08248 b1 AD-20578- 751 1309 1821 0.061816 0.0668734790.00043 b1 AD-20579- 752 1310 1822 0.264159 0.12924691 0.10334 b1AD-20581- 754 1311 1823 0.513809 0.48572165 b1 AD-20582- 755 1312 18240.389395 0.399959136 b1 AD-20625- 756 1313 1825 0.256936 0.1164334890.01077 b1 AD-20626- 757 1314 1826 0.087123 0.081492545 0.00028 b1AD-20627- 758 1315 1827 0.055328 0.055650353 b1 AD-20628- 759 1316 18280.447781 0.303068664 b1 AD-20629- 760 1317 1829 0.234948 0.307299345 b1AD-20630- 761 1318 1830 0.489721 0.223370205 b1 AD-20631- 762 1319 18310.653686 0.400035988 b1 AD-20632- 763 1320 1832 0.770776 0.563518413 b1AD-20633- 781 1321 1833 0.19814 0.117243348 0.04183 b1 AD-20634- 7991322 1834 0.112208 0.080929635 0.0041 b1 AD-20635- 800 1323 18350.415981 0.277900171 b1 AD-20636- 801 1324 1836 0.935782 0.977777409 b1AD-20637- 802 1325 1837 0.867293 0.869090293 b1 AD-20638- 803 1326 18380.26502 0.374396441 b1 AD-20639- 804 1327 1839 0.283056 0.375662409 b1AD-20640- 805 1328 1840 0.057288 0.069780308 0.00527 b1 AD-20641- 8061329 1841 0.533881 0.623267984 b1 AD-20642- 807 1330 1842 0.4600050.389433419 b1 AD-20643- 808 1331 1843 0.315391 0.221106278 b1 AD-20644-809 1332 1844 0.109044 0.131007072 0.00111 b1 AD-20645- 810 1333 18451.128482 0.998274628 b1 AD-20646- 811 1334 1846 0.095785 0.0668734790.00008 b1 AD-20647- 812 1335 1847 1.156048 0.143850149 b1 AD-20648- 8131336 1848 0.106528 0.109039633 0.00115 b1 AD-20649- 814 1337 18490.706715 1.084879386 b1 AD-20650- 815 1338 1850 0.171848 0.2557341260.02769 b1 AD-20651- 816 1339 1851 0.611334 0.869126829 b1 AD-20652- 8171340 1852 0.143195 0.207349809 0.00003 b2 AD-20653- 818 1341 18530.193334 0.079856194 0.00532 b1 AD-20654- 819 1342 1854 0.755821.143572737 b1 AD-20655- 820 1343 1855 0.858399 1.313052418 b1 AD-20656-821 1344 1856 0.541334 0.428549767 b1 AD-20657- 822 1345 1857 0.7419040.863123331 b1 AD-20658- 823 1346 1858 0.198178 0.475597123 b1 AD-20659-824 1347 1859 0.388124 0.491623427 b1 AD-20660- 825 1348 1860 0.0774390.077627767 0.00955 b1 AD-20661- 826 1349 1861 0.156255 0.1375128770.28191 b1 AD-20662- 847 1351 1863 0.476044 0.505032662 b1 AD-20663- 8491353 1865 0.600475 0.625882515 b1 AD-20664- 850 1354 1866 0.6525881.119554306 b1 AD-20665- 851 1355 1867 0.714432 1.029952624 b1 AD-20666-852 1356 1868 0.864059 1.029903142 b1 AD-20667- 853 1357 1869 0.9133261.426528593 b1 AD-20668- 854 1358 1870 1.060234 1.57734692 b1 AD-20669-855 1359 1871 1.067968 1.588470861 b1 AD-20670- 856 1360 1872 0.5344460.427490356 b1 AD-20671- 857 1361 1873 0.134052 0.089804062 0.02226 b1AD-20672- 858 1362 1874 0.256251 0.37063074 b1 AD-20673- 859 1363 18750.768658 1.062935788 b1 AD-20674- 860 1364 1876 0.619496 0.691770914 b1AD-20675- 861 1365 1877 1.067506 1.213141054 b1 AD-20676- 862 1366 18780.359737 0.305314129 b1 AD-20677- 915 1373 1885 0.870382 1.317223358 b1AD-20678- 965 1385 1897 0.326325 0.270354544 b1 AD-20679- 966 1386 18980.962322 1.769753135 b1 AD-20680- 967 1387 1899 0.948845 1.697660683 b1AD-20681- 968 1388 1900 0.798916 0.875277199 b1 AD-20682- 969 1389 19011.087386 1.561791318 b1 AD-20683- 970 1390 1902 1.110234 1.477899464 b1AD-20684- 971 1391 1903 0.882193 1.207906399 b1 AD-20685- 972 1392 19040.8211 1.143298164 b1 AD-20686- 973 1393 1905 0.789661 1.092425319 b1AD-20687- 974 1394 1906 0.768455 1.089410843 b1 AD-20688- 975 1395 19070.770702 1.022838222 b1 AD-20689- 976 1396 1908 0.771035 1.378134406 b1AD-20690- 977 1397 1909 0.823135 1.037066685 b1 AD-20691- 978 1398 19100.831763 1.326512826 b1 AD-20692- 979 1399 1911 0.972934 1.15075282 b1AD-20693- 1011 1400 1912 0.072624 0.091035811 0.00046 b1 AD-20694- 10121401 1913 0.066949 0.06070901 0.00114 b1 AD-20695- 1013 1402 19140.749807 0.933655977 b1 AD-20696- 1014 1403 1915 1.000287 1.346484427 b1AD-20697- 1015 1404 1916 0.809563 1.359618312 b1 AD-20698- 1016 14051917 0.592378 0.887325083 b1 AD-20699- 1048 1406 1918 0.5549440.857161303 b1 AD-20700- 1049 1407 1919 0.105695 0.123924613 0.04345 b1AD-20701- 1050 1408 1920 0.650858 0.845462044 b1 AD-20702- 1051 14091921 0.046369 0.069008817 0.0001 b1 AD-20703- 1053 1411 1923 0.3964510.459473782 b1 AD-20704- 1054 1412 1924 0.812095 0.881195881 b1AD-20705- 1055 1413 1925 0.798647 0.971064227 b1 AD-20706- 1329 14161928 0.405034 0.323806017 b1 AD-20707- 1330 1417 1929 0.043570.060698804 <1E−05 b1 AD-20709- 1332 1418 1930 0.13188 0.1205445750.66004 b1 AD-20710- 1333 1419 1931 0.401696 0.366834618 0.00717 b1AD-20711- 1334 1420 1932 0.43653 0.442183347 b1 AD-20712- 1335 1421 19330.584644 0.928468269 b1 AD-20713- 1336 1422 1934 0.289461 0.287688205 b1AD-20714- 1337 1423 1935 0.088346 0.060267944 0.01035 b1 AD-20715- 13381424 1936 0.741225 0.630689818 b1 AD-20716- 1339 1425 1937 0.073550.074476264 0.00203 b1 AD-20717- 1359 1426 1938 0.873116 0.931417569 b1AD-20718- 1360 1427 1939 0.399209 0.291704183 b1 AD-20719- 1361 14281940 0.887602 0.831002378 b1 AD-20720- 1362 1429 1941 0.6360850.563508261 b1 AD-20721- 1363 1430 1942 0.844185 0.872317061 b1AD-20722- 1364 1431 1943 0.862113 1.431782046 b1 AD-20723- 1365 14321944 0.842047 0.991468351 b1 AD-20724- 1366 1433 1945 0.7156330.884239241 b1 AD-20725- 1367 1434 1946 0.8211 1.13103112 b1 AD-20726-1368 1435 1947 0.870382 0.99848447 b1 AD-20727- 1369 1436 1948 0.8397220.869231218 b1 AD-20728- 1370 1437 1949 0.146011 0.171134401 0.20951 b1AD-20729- 1371 1438 1950 0.842855 0.720731925 b1 AD-20730- 1372 14391951 0.052245 0.060685683 0.00339 b1 AD-20731- 1373 1440 1952 0.7195740.38625851 b1 AD-20732- 1374 1441 1953 1.386744 1.054026625 b1 AD-20733-1375 1442 1954 1.396389 1.27991671 b1 AD-20734- 1379 1446 1958 1.2946241.275419601 b1 AD-20735- 1380 1447 1959 1.209551 1.219180143 b1AD-20736- 1381 1448 1960 1.238102 1.297970725 b1 AD-20737- 1382 14491961 1.294966 1.241884392 b1 AD-20738- 1383 1450 1962 1.1308191.018122609 b1 AD-20739- 1384 1451 1963 1.166108 1.181930519 b1AD-20740- 1385 1452 1964 1.178614 0.940155929 b1 AD-20741- 1386 14531965 0.079687 0.065054016 0.00064 b1 AD-20742- 1387 1454 1966 1.3915331.415064673 b1 AD-20743- 1407 1455 1967 0.29152 0.304315297 b1 AD-20744-1408 1456 1968 0.908655 0.79362811 b1 AD-20745- 1409 1457 1969 1.4464671.335038609 b1 AD-20746- 1410 1458 1970 1.170192 0.719886814 b1AD-20747- 1411 1459 1971 1.432098 1.21912157 b1 AD-20748- 1428 1460 19720.772113 0.538061131 b1 AD-20749- 1429 1461 1973 1.102275 0.893650074 b1AD-20750- 1430 1462 1974 1.194895 0.711852844 b1 AD-20751- 1431 14631975 0.667424 0.418146571 b1 AD-20752- 1432 1464 1976 1.3915331.266609634 b1 AD-20753- 1433 1465 1977 1.417073 0.853311991 b1AD-20754- 1434 1466 1978 0.844918 0.57890069 b1 AD-20755- 1435 1467 19791.416154 1.696464074 b1 AD-20756- 1436 1468 1980 1.836139 1.028732572 b1AD-20757- 1437 1469 1981 1.38232 1.06207192 b1 AD-20758- 1438 1470 19821.330733 1.266974756 b1 AD-20759- 1439 1471 1983 1.451044 1.325816823 b1AD-20760- 1440 1472 1984 1.130602 0.940268849 b1 AD-20761- 1441 14731985 1.095795 0.976601567 b1 AD-20762- 1442 1474 1986 1.2033791.068688943 b1 AD-20763- 1443 1475 1987 1.001156 1.036261261 b1AD-20764- 1444 1476 1988 0.118576 0.069158478 0.01035 b1 AD-20765- 14451477 1989 1.280472 0.649176427 b1 AD-20766- 1446 1478 1990 0.8598880.413525996 b1 AD-20767- 1447 1479 1991 0.915437 0.874208876 b1AD-20768- 1448 1480 1992 1.491623 1.033674481 b1 AD-20769- 1449 14811993 1.14688 0.779689688 b1 AD-20770- 1450 1482 1994 1.0260430.755677199 b1 AD-20771- 1451 1483 1995 1.220002 1.129786315 b1AD-20772- 1452 1484 1996 1.055092 1.018073695 b1 AD-20773- 1453 14851997 1.170473 1.035863089 b1 AD-20774- 1454 1486 1998 0.8484290.63553784 b1 AD-20775- 1455 1487 1999 1.025895 0.62466094 b1 AD-20776-1456 1488 2000 1.114715 0.89556372 b1 AD-20777- 1457 1489 2001 1.1071750.712453613 b1 AD-20778- 1458 1490 2002 1.138466 1.114392802 b1AD-20779- 1459 1491 2003 1.377165 0.815548171 b1 AD-20780- 1460 14922004 1.595425 1.004202298 b1 AD-20781- 1461 1493 2005 1.2498941.014642825 b1 AD-20782- 1462 1494 2006 1.130439 0.697841818 b1AD-20783- 1482 1495 2007 0.313756 0.140716947 0.07259 b1 AD-20784- 14831496 2008 0.372852 0.269030566 b1 AD-20785- 1484 1497 2009 0.3431430.300598832 b1 AD-20786- 1485 1498 2010 0.921805 0.835570424 b1AD-20787- 1486 1499 2011 1.091775 0.749006565 b1 AD-20788- 1487 15002012 1.080502 1.028732572 b1 AD-20789- 1547 1515 2027 0.8245630.771592653 b1 AD-20790- 1548 1516 2028 0.497168 0.351347846 b1AD-20791- 1549 1517 2029 1.37743 1.386042945 b1 AD-20792- 1602 1518 20301.812166 0.920719325 b1 AD-20793- 1603 1519 2031 1.643864 1.253455962 b1AD-20794- 1604 1520 2032 1.406609 1.102568271 b1 AD-20795- 1605 15212033 1.25418 0.893349812 b1 AD-20796- 1606 1522 2034 1.2082461.054279832 b1 AD-20797- 1634 1526 2038 1.182812 1.134873015 b1AD-20798- 1635 1527 2039 0.816019 0.877112215 b1 AD-20799- 1636 15282040 1.637852 0.777034161 b1 AD-20800- 1699 1530 3282 1.267221.137480645 b1 AD-20801- 1700 1531 3283 0.690961 0.517931545 b1AD-20868- 848 1352 1864 1.043896 0.644316747 b1 AD-20869- 1052 1410 19220.759026 0.601255433 b1 AD-20870- 1607 1523 2035 0.963852 0.693770729 b1AD-20871- 1608 1524 2036 1.440675 1.47009696 b1 AD-20872- 1633 1525 20370.988535 0.618366913 b1 AD-20873- 1698 1529 2041 1.506987 0.994836178 b1*wherein <1E−05 means less than 1 × 10⁻⁵

Example 3A Methodology for In Vitro Screening

Cell Culture and Transfections

WI-38 or HeLa (ATCC, Manassas, Va.) cells are grown to near confluenceat 37° C. in an atmosphere of 5% CO₂ in RPMI or EMEM (ATCC) supplementedwith 10% FBS, streptomycin, and glutamine (ATCC) before being releasedfrom the plate by trypsinization. Reverse transfection is carried out byadding 5 μl of Opti-MEM to 5 μl of siRNA duplexes per well into a96-well plate along with 10 μl of Opti-MEM plus 0.2 μl of LipofectamineRNAiMax per well (Invitrogen, Carlsbad Calif. cat #13778-150) andincubated at room temperature for 15 minutes. 80 μl of complete growthmedia without antibiotic containing 2×10⁴ WI-38 cells or 2.0×10⁴ Helacells are then added. Cells are incubated for 24 hours prior to RNApurification. Experiments are performed l OnM final duplex concentrationfor WI-38 cells and at 10 nM and 0.1 nM final duplex concentration forHeLa cells for each of the HSF1 siRNAs. A subset of siRNAs that showedrobust silencing in the 10 nM and 0.1 nM screens are assayed over arange of concentrations from 10 nM to 0.00005M to determine their IC50.

Total RNA isolation using MagMAX-96 Total RNA Isolation Kit (AppliedBiosystem, Foster City Calif., part #: AM1830).

Cells are harvested and lysed in 140 μl of Lysis/Binding Solution thenmixed for 1 minute at 850 rpm using an Eppendorf Thermomixer (the mixingspeed is the same throughout the process). Twenty microliters ofmagnetic beads and Lysis/Binding Enhancer mixture are added intocell-lysate and mixed for 5 minutes. Magnetic beads are captured usingmagnetic stand and the supernatant is removed without disturbing thebeads. After removal of the supernatant, magnetic beads are washed withWash Solution 1 (isopropanol added) and mixed for 1 minute. Beads arecaptured again and supernatant removed. Beads are then washed with 150μl Wash Solution 2 (Ethanol added), captured and supernatant is removed.50 ul of DNase mixture (MagMax turbo DNase Buffer and Turbo DNase) isthen added to the beads and they are mixed for 10 to 15 minutes. Aftermixing, 100 μl of RNA Rebinding Solution is added and mixed for 3minutes. Supernatant is removed and magnetic beads are washed again with150 μl Wash Solution 2 and mixed for 1 minute and supernatant is removedcompletely. The magnetic beads are mixed for 2 minutes to dry before RNAis eluted with 50 μl of water.

cDNA synthesis is performed using ABI High capacity cDNA reversetranscription kit (Applied Biosystems, Cat #4368813) as follows: Amaster mix of 2 μl 10× Buffer, 0.8 μl 25×dNTPs, 2 μl Random primers, 1μl Reverse Transcriptase, 1 μl RNase inhibitor and 3.2 μl of H₂O perreaction are added into 10 μl total RNA. cDNA is generated using aBio-Rad C-1000 or S-1000 thermal cycler (Hercules, Calif.) through thefollowing steps: 25° C. 10 min, 37° C. 120 min, 85° C. 5 sec, 4° C.hold.

Real time PCR is performed as follows: 2 μl of cDNA are added to amaster mix containing 0.5 ul GAPDH TaqMan Probe (Applied Biosystems Cat#4326317E) or 18S TaqMan Probe (Applied Biosystems Cat #4319413E), 0.5μl HSF1 TaqMan probe (Applied Biosystems cat #HS00232134 M1) and 5 μlRoche Probes Master Mix (Roche Cat #04887301001) in a total of 10 μl perwell in a LightCycler 480 384 well plate (Roche cat #0472974001). Realtime PCR is done in a LightCycler 480 Real Time PCR machine (Roche).Each duplex is tested in at least two independent transfections. Eachtransfection is assayed in duplicate.

Real time data are analyzed using the ΔΔCt method (Livak et al 2001).Each sample is normalized to GAPDH expression and knockdown is assessedrelative to cells transfected with the non-targeting duplex AD-1955.IC50s are defined using a 4 parameter fit model in XLfit. See e.g.,Kenneth and Schmittge. Methods 25:4, December 2001, 402-408.

Example 4 HSF1 siRNAs

As shown in Table 5, a subset of 101 HSF1 RNAi agents is presented.

Many of the 101 selected duplexes had >80% gene knockdown (KD) in bothHeLa and WI-38 cell lines at 10 nM. Also included in the 101 areduplexes that had >80% KD in a single cell line, provided that theaverage KD of the two lines is >75% KD. This set also includes allduplexes that have >50% KD in WI-38 in the 0.1nM screen. These 101 HSF1siRNAs of particular interest are shown in Table 2. This table alsopresents the knockdown (KD) in HeLa and WI-38 cells when 10 nM or 0.1 nMof siRNA was used. 1.000=100% gene expression, or no or 0% geneknockdown; and 0.000=0% gene expression, or complete or 100% geneknockdown. For example, for AD-20303, the “HeLa 10 nM” column indicates“0.099”, meaning that at this concentration in these cells, the RNAiagent reduced gene expression to 9.9%, or exhibited 90.1% geneknockdown. In WI-38 cells at a concentration of 10 nM, this RNAi agentexhibited 5.6% residual gene activity, or 94.4% gene knockdown.

TABLE 5 HSF1 RNAi agents - Knock Down Data. Average WI-38 Duplex HeLa 10nM WI-38 10 nM (10 nM) 0.1 nM 1 AD-20303 0.099 0.056 0.078 0.529 2AD-20313 0.127 0.090 0.109 0.884 3 AD-20315 0.221 0.188 0.205 0.759 4AD-20348-b1 0.173 0.111 0.142 0.805 5 AD-20362-b1 0.253 0.201 0.2270.759 6 AD-20364-b1 0.231 0.164 0.198 0.903 7 AD-20365-b1 0.128 0.1560.142 0.738 8 AD-20366-b1 0.173 0.207 0.190 0.802 9 AD-20373-b1 0.1050.098 0.101 0.825 10 AD-20376-b1 0.067 0.051 0.059 0.656 11 AD-20377-b10.207 0.143 0.175 0.848 12 AD-20386-b1 0.061 0.030 0.046 0.379 13AD-20389-b1 0.082 0.067 0.075 0.802 14 AD-20391-b1 0.064 0.032 0.0480.415 15 AD-20392-b1 0.093 0.032 0.062 0.448 16 AD-20397-b1 0.167 0.1100.138 0.862 17 AD-20398-b1 0.229 0.139 0.184 0.998 18 AD-20399-b1 0.0710.067 0.069 0.658 19 AD-20401-b1 0.079 0.072 0.075 0.883 20 AD-20402-b10.067 0.047 0.057 0.476 21 AD-20403-b1 0.041 0.034 0.037 0.462 22AD-20404-b1 0.095 0.060 0.077 0.695 23 AD-20406-b1 0.262 0.105 0.1830.655 24 AD-20407-b1 0.123 0.047 0.085 0.589 25 AD-20408-b1 0.100 0.0430.071 0.540 26 AD-20409-b1 0.183 0.095 0.139 0.653 27 AD-20410-b1 0.0930.073 0.083 0.748 28 AD-20411-b1 0.139 0.081 0.110 0.669 29 AD-20413-b10.189 0.124 0.156 0.687 30 AD-20422-b1 0.093 0.060 0.077 0.392 31AD-20428-b1 0.289 0.132 0.210 0.865 32 AD-20434-b1 0.129 0.076 0.1030.547 33 AD-20435-b1 0.072 0.040 0.056 0.375 34 AD-20437-b1 0.064 0.0320.048 0.245 35 AD-20438-b1 0.060 0.027 0.043 0.477 36 AD-20439-b1 0.1140.086 0.100 0.387 37 AD-20487-b1 0.060 0.024 0.042 0.122 38 AD-20488-b10.070 0.036 0.053 0.480 39 AD-20489-b1 0.057 0.030 0.044 0.205 40AD-20490-b1 0.087 0.033 0.060 0.308 41 AD-20491-b1 0.077 0.035 0.0560.259 42 AD-20493-b1 0.095 0.037 0.066 0.512 43 AD-20495-b1 0.095 0.0730.084 0.690 44 AD-20502-b1 0.256 0.085 0.171 0.753 45 AD-20507-b1 0.0700.033 0.051 0.250 46 AD-20513-b1 0.152 0.074 0.113 0.804 47 AD-20527-b10.228 0.188 0.208 0.861 48 AD-20535-b1 0.149 0.175 0.162 1.034 49AD-20544-b1 0.053 0.175 0.114 0.955 50 AD-20545-b1 0.047 0.072 0.0590.667 51 AD-20546-b1 0.147 0.168 0.158 0.840 52 AD-20547-b1 0.135 0.1920.164 0.886 53 AD-20548-b1 0.040 0.041 0.040 0.383 54 AD-20549-b1 0.1440.199 0.171 0.864 55 AD-20552-b1 0.129 0.139 0.134 0.790 56 AD-20555-b10.127 0.122 0.124 0.709 57 AD-20556-b1 0.071 0.159 0.115 0.786 58AD-20557-b1 0.046 0.077 0.062 0.678 59 AD-20558-b1 0.094 0.157 0.1250.840 60 AD-20560-b1 0.036 0.035 0.036 0.295 61 AD-20561-b1 0.077 0.0990.088 0.752 62 AD-20562-b1 0.042 0.040 0.041 0.556 63 AD-20563-b1 0.0270.028 0.027 0.471 64 AD-20564-b1 0.029 0.028 0.028 0.222 65 AD-20565-b10.055 0.061 0.058 0.547 66 AD-20566-b1 0.027 0.043 0.035 0.598 67AD-20572-b1 0.048 0.077 0.062 0.855 68 AD-20574-b1 0.203 0.228 0.2160.926 69 AD-20575-b1 0.102 0.125 0.113 0.907 70 AD-20577-b1 0.133 0.2240.178 0.837 71 AD-20578-b1 0.067 0.062 0.064 0.371 72 AD-20579-b1 0.1290.264 0.197 0.826 73 AD-20625-b1 0.116 0.257 0.187 0.942 74 AD-20626-b10.081 0.087 0.084 0.929 75 AD-20627-b1 0.056 0.055 0.055 0.698 76AD-20633-b1 0.117 0.198 0.158 0.882 77 AD-20634-b1 0.081 0.112 0.0970.955 78 AD-20640-b1 0.070 0.057 0.064 0.972 79 AD-20644-b1 0.131 0.1090.120 0.590 80 AD-20646-b1 0.067 0.096 0.081 0.739 81 AD-20648-b1 0.1090.107 0.108 0.906 82 AD-20650-b1 0.256 0.172 0.214 1.204 83 AD-20652-b10.037 0.029 0.033 0.371 84 AD-20653-b1 0.080 0.193 0.137 1.286 85AD-20660-b1 0.078 0.077 0.078 1.097 86 AD-20661-b1 0.138 0.156 0.1471.368 87 AD-20671-b1 0.090 0.134 0.112 1.127 88 AD-20693-b1 0.091 0.0730.082 0.515 89 AD-20694-b1 0.061 0.067 0.064 0.633 90 AD-20700-b1 0.1240.106 0.115 0.890 91 AD-20702-b1 0.069 0.046 0.058 0.864 92 AD-20707-b10.061 0.044 0.052 0.519 93 AD-20708-b1 0.217 0.182 0.199 1.529 94AD-20709-b1 0.121 0.132 0.126 0.736 95 AD-20714-b1 0.060 0.088 0.0741.221 96 AD-20716-b1 0.074 0.074 0.074 0.697 97 AD-20728-b1 0.171 0.1460.159 0.794 98 AD-20730-b1 0.061 0.052 0.056 0.484 99 AD-20741-b1 0.0650.056 0.061 0.367 100 AD-20764-b1 0.069 0.083 0.076 0.510 101AD-20783-b1 0.141 0.221 0.181 0.603

Example 5 HeLa Cell Screen of HSF1 RNAi Agents

The 101 duplexes in Table 2 were screened in HeLa cells in a doseresponse screen. The purpose of this screen was to determine the EC50(minimum dosage of RNAi agent capable of reducing gene expression by50%).

A serial dilution was used, comprising 12 concentrations between 10 nMand 1×10⁻⁵ nM. The siRNA duplexes were transfected into cells usingLipofectamine RNAiMax (Zhao et al. 2008 Mol. Biotech. 40: 19-26). After24 hours total RNA is isolated and used for cDNA synthesis using randomprimers. 10-20 ng of total cDNA is used for TaqMan assays (AppliedBiosystems, Foster City, Calif.). Each duplex was assayed in duplicate.Data are expressed as the average-fold change compared to thenon-targeting control AD-1955, which does not bind to HSF1. AD1955 is asiRNA targeting firefly luciferase and is used as a negative controlsiRNA in screening assays. The results are shown in Table 6.

TABLE 6 EC50 data* EC50 Average EC50 Duplex ID EC50 (nM) EC50 (nM) (nM)(nM) AD-20303-b1 0.06064 0.01286 0.03675 AD-20313-b1 0.22911 0.079290.15420 AD-20315-b1 0.00920 0.00358 0.00639 AD-20348-b1 1.12593 0.449380.78766 AD-20362-b1 0.88761 0.83959 0.86360 AD-20364-b1 0.05833 0.090570.07445 AD-20365-b1 0.01115 0.01565 0.01340 AD-20366-b1 0.25818 0.273110.26565 AD-20373-b1 0.01657 0.03293 0.02475 AD-20376-b1 0.00400 0.006140.00507 AD-20377-b1 0.46977 0.75572 0.61274 AD-20386-b1 0.00054 0.000450.00049 AD-20389-b1 0.08152 0.07000 0.07576 AD-20391-b1 0.00569 0.003570.00463 AD-20392-b1 0.02855 0.05939 0.04397 AD-20397-b1 0.17323 0.291370.23230 AD-20398-b1 0.54902 0.41237 0.48070 AD-20399-b1 0.02232 0.012810.01757 AD-20401-b1 0.08227 0.08691 0.08459 AD-20402-b1 0.00902 0.005080.00705 AD-20403-b1 0.00110 0.00246 0.00178 AD-20404-b1 0.00850 0.009120.00881 AD-20406-b1 0.00842 0.00914 0.00878 AD-20407-b1 0.00236 0.001300.00183 AD-20408-b1 0.00341 0.00289 0.00315 AD-20409-b1 0.00396 0.002190.00307 AD-20410-b1 0.01225 0.01775 0.01500 AD-20411-b1 0.00283 0.001040.00193 AD-20413-b1 No EC50 No EC50 AD-20422-b1 0.00160 0.00155 0.00157AD-20428-b1 1.37427 1.57813 1.47620 AD-20434-b1 0.00212 0.00456 0.00334AD-20435-b1 0.00040 0.00032 0.00036 AD-20437-b1 <1E−05 <1E−05AD-20438-b1 0.00006 0.00007 0.00007 AD-20439-b1 0.00029 0.00010 0.00020AD-20487-b1 <1E−05 <1E−05 AD-20488-b1 0.00020 0.00013 0.00016AD-20489-b1 <1E−05 <1E−05 AD-20490-b1 0.00026 0.00015 0.00021AD-20491-b1 0.00004 <1E−05 0.00004 AD-20493-b1 0.00218 0.00173 0.00196AD-20495-b1 0.03348 0.00792 0.0168299 0.01941 AD-20502-b1 0.039100.00037 0.0125045 0.01732 AD-20507-b1 >10 0.22784 >10 0.22784AD-20513-b1 0.09514 <1E−05 0.0329948 0.06407 AD-20527-b1 0.08362 0.038720.0415847 0.05464 AD-20535-b1 0.58303 0.00181 0.1768769 0.25390AD-20544-b1 0.01126 0.00062 0.1768769 0.06292 AD-20545-b1 0.011930.00229 0.00711 AD-20546-b1 0.36163 0.04777 0.20470 AD-20547-b1 0.058300.01273 0.03551 AD-20548-b1 0.00108 0.00013 0.00061 AD-20549-b1 0.250580.02762 0.13910 AD-20552-b1 0.13845 0.03697 0.08771 AD-20555-b1 0.028630.01263 0.02063 AD-20556-b1 0.01001 0.00508 0.0463849 0.02049AD-20557-b1 0.00091 0.00073 0.0085311 0.00339 AD-20558-b1 0.079060.06566 0.2598412 0.13485 AD-20560-b1 <1E−05 <1E−05 <1E−05 AD-20561-b10.02092 0.02674 0.0519951 0.03322 AD-20562-b1 0.00051 0.00046 0.00220010.00105 AD-20563-b1 0.00004 0.00003 0.0006466 0.00024 AD-20564-b1 <1E−05<1E−05 <1E−05 AD-20565-b1 0.01335 0.00991 0.0159182 0.01306 AD-20566-b10.00296 0.00239 0.0030765 0.00281 AD-20572-b1 0.06851 0.05459 0.26569890.12960 AD-20574-b1 0.86331 0.70143 2.5513791 1.37204 AD-20575-b10.13577 0.13435 0.2985167 0.18954 AD-20577-b1 0.07983 0.08513 0.19310930.11936 AD-20578-b1 0.00050 0.00035 0.00043 AD-20579-b1 0.12105 0.085620.10334 AD-20625-b1 0.01117 0.01037 0.01077 AD-20626-b1 0.00030 0.000270.00028 AD-20627-b1 <1E−05 <1E−05 AD-20633-b1 0.04183 0.04183 0.04183AD-20634-b1 0.00371 0.00450 0.00410 AD-20640-b1 0.00760 0.00294 0.00527AD-20644-b1 0.00098 0.00123 0.00111 AD-20646-b1 0.00007 0.00008 0.00008AD-20648-b1 0.00104 0.00126 0.00115 AD-20650-b1 0.00059 0.05479 0.02769AD-20652-b1 0.00003 <1E−05 0.00003 AD-20653-b1 0.00887 0.00177 0.00532AD-20660-b1 0.01515 0.00394 0.00955 AD-20661-b1 0.45059 0.11322 0.28191AD-20671-b1 0.02504 0.01948 0.02226 AD-20693-b1 0.00080 0.00013 0.00046AD-20694-b1 0.00105 0.00123 0.00114 AD-20700-b1 0.04602 0.04089 0.04345AD-20702-b1 0.00015 0.00006 0.00010 AD-20707-b1 <1E−05 <1E−05AD-20708-b1 0.99910 0.32099 0.66004 AD-20709-b1 0.01314 0.00120 0.00717AD-20714-b1 0.00599 0.01470 0.01035 AD-20716-b1 0.00303 0.00103 0.00203AD-20728-b1 0.26241 0.15661 0.20951 AD-20730-b1 0.00331 0.00346 0.00339AD-20741-b1 0.00094 0.00033 0.00064 AD-20764-b1 0.01432 0.00638 0.01035AD-20783-b1 0.08658 0.05859 0.07259 *wherein “<1E−05” means less than 1× 10⁻⁵.

Example 6 Additional Screening of HSF1 RNAi Agents

After making the determinations reported in Tables 4 and 5, it wasdetermined that certain of the oligonucleotides were subject toconcentration artifacts now thought to arise from incomplete thawing ofplates containing the oligonucleotides. Subsequent analysis of two largedatasets (unrelated to Target) indicated that any such concentrationartifacts would not alter the assay results more than that which can beaccounted for by normal experimental variability. Moreover, theseconcentration artifacts are unlikely to substantially affect ourconclusions about lead selection.

Additional screening data of the HSF1 siRNAs described above ispresented in Table 7, below.

HSF1 rescreen

HSF1 siRNAs are rescreened to address discrepancies that are found induplex concentrations when the single strands for these duplexes areoriginally annealed. 414 siRNAs are reannealed and screened at two siRNAconcentrations, 10 nM and 0.1 nM. All duplexes are screened at bothdoses in two independent experiments.

In Vitro Screening:

Cell Culture and Transfections.

HeLa (ATCC, Manassas, Va.) cells are grown to near confluence at 37° C.in an atmosphere of 5% CO₂ in EMEM (ATCC) supplemented with 10% FBS,streptomycin, and glutamine (ATCC) before being released from the plateby trypsinization. Reverse transfection is carried out by adding to 5 μlof 200 nM or 2 nM siRNA duplex per well into a 96-well plate along with15 μl of Opti-MEM plus 0.2 μl of Lipofectamine RNAiMax per well(Invitrogen, Carlsbad Calif. cat #13778-150) and incubated at roomtemperature for 15 minutes. 80 μl of complete growth media withoutantibiotic containing 2×10⁴ Hela cells are then added resulting in afinal duplex concentration of 10 nM or 0.1 nM. Cells are incubated for24 hours prior to RNA purification.

mRNA Isolation Using Dynobeads (Invitrogen, Carlsbad Calif. cat#610-12):

Cells are harvested and lysed in 150 μl of lysis buffer then mixed for 5minutes at 850 rpm using and platform shaker (the mixing speed is thesame throughout the process). Ten micro liters of magnetic beads thathad previously been washed in 70 ul of lysis buffer are added intocell-lysate and mixed for 5 minutes. Magnetic beads are captured usingmagnetic stand and the supernatant is removed without disturbing thebeads. Magnetic beads/RNA are washed twice with wash buffer A by adding150 ul of buffer, shaking for 1 minute and discarding supernatantfollowing capture of beads on magnetic stand. Beads are then washed with150 μl wash buffer B, agitated for 1 minute, captured a magnetic beadstand and supernatant is removed. The Dynobead/mRNA mixture is thenwashed with 150u1 of elution buffer by shaking for 1 minute, capturingon a magnetic ring stand and discarding the elution buffer. mRNA iseluted by adding 50 ul of elution buffer and shaking for 4 minutes at70° C. After mixing plates are placed on the magnetic bead stand for 2minutes to recapture the beads before the supernatant containing theeluted RNA is removed to a new plate.

cDNA Synthesis Using ABI High Capacity cDNA Reverse Transcription Kit

A master mix of 1 μl 10× Buffer, 0.41125×dNTPs, 1 μl Random primers, 0.5μl Reverse Transcriptase, 0.5 μl RNase inhibitor and 1.6 μl of H₂O perreaction are added into 5 μl total RNA. cDNA is generated using aBio-Rad C-1000 or S-1000 thermal cycler (Hercules, Calif.) through thefollowing steps: 25° C. 10 min, 37° C. 120 min, 85° C. 5 sec, 4° C.hold.

Real time PCR: 2 μl of cDNA are added to a master mix containing 0.5 ulGAPDH TaqMan Probe (Applied Biosystems Cat #4326317E) or 18S TaqManProbe (Applied Biosystems Cat #4319413E), 0.5 μl HSF1 TaqMan probe(Applied Biosystems cat #HS00232134 M1) and 5 μl Roche Probes Master Mix(Roche Cat #04887301001) in a total of 10 μl per well in a LightCycler480 384 well plate (Roche cat #0472974001). Real time PCR is done in aLightCycler 480 Real Time PCR machine (Roche). Each duplex is tested inat two independent transfections (Called Screen 1 and Screen 2). Eachtransfection is assayed in duplicate. For Screen 1 and Screen 2, qPCR isperformed using GAPDH for normalization. For Screen 1, qPCR is repeateda second time using 18S for normalization.

Real time data are analyzed using the ΔΔCt method (Livak et al 2001).Each sample is normalized to GAPDH or 18S expression and knockdown isassessed relative to cells transfected with the non-targeting duplexAD-1955. Higher values represent less knockdown. Reference: Analysis ofRelative Gene Expression Data Using Real-Time Quantitative PCR and the2-ΔΔCT Method. Kenneth J. Livak, Thomas D. Schmittge. Methods 25:4,December 2001, 402-408.

As shown in Table 7, below, certain HSF1 siRNAs are re-screened twice(Screens 1 and 2), using GAPDH and 18s as controls. This table alsoincludes screening data for several HSF1 siRNAs without data presentedabove. Numbers represent residual HSF1 gene activity, with GAPDH or 18sas control reference genes. Thus, for example, for AD-20278-b2, “0.08”at “10 nM Screen 1 (GAPDH)” indicates 8% residual HSF1 gene activity, or92% gene knockdown.

TABLE 7 ADDITIONAL SCREENING OF HSF1 siRNAs (10 nM and 0.1 nM) 10 nM 10nM 10 nM 0.1 nM 0.1 nM 0.1 nM Screen 1 Screen 1 Screen 2 Screen 1 Screen1 Screen 2 Duplex ID (GAPDH) (18s) (GAPDH) (GAPDH) (18s) (GAPDH)AD-20278-b2 0.08 0.07 0.05 0.49 0.15 0.39 AD-20279-b2 0.06 0.06 0.050.26 0.20 0.27 AD-20280-b2 0.19 0.21 0.15 0.71 0.57 0.70 AD-20281-b20.11 0.07 0.08 0.56 0.48 0.54 AD-20282-b2 0.10 0.08 0.05 1.13 0.89 1.13AD-20283-b2 0.11 0.10 0.08 1.10 1.05 1.06 AD-20284-b2 0.78 0.94 0.711.86 0.92 1.12 AD-20285-b2 0.83 1.18 1.11 1.19 0.96 1.08 AD-20286-b21.04 1.20 0.97 1.20 0.96 1.08 AD-20287-b2 1.06 1.30 1.06 1.03 1.04 1.05AD-20288-b2 0.89 0.70 0.95 0.98 1.01 1.07 AD-20289-b2 1.42 1.43 1.381.24 1.07 1.02 AD-20290-b2 1.05 0.95 1.09 1.03 0.92 1.05 AD-20291-b20.98 0.86 0.91 1.29 0.92 1.12 AD-20292-b2 1.04 0.93 1.06 1.16 1.05 1.18AD-20293-b2 0.90 0.81 0.95 1.11 0.82 1.16 AD-20294-b2 0.81 0.71 0.771.22 0.95 1.21 AD-20295-b2 0.85 0.90 0.86 0.97 0.85 1.00 AD-20296-b20.34 0.27 0.30 1.02 0.80 1.09 AD-20297-b2 0.78 0.73 0.86 0.99 0.82 1.01AD-20298-b2 1.65 0.77 0.89 1.24 1.03 1.05 AD-20299-b2 0.88 0.81 0.911.12 0.95 1.05 AD-20300-b2 0.83 0.52 0.75 1.12 0.83 1.07 AD-20301-b20.99 0.42 0.81 0.88 0.99 0.85 AD-20302-b2 0.97 1.14 1.04 0.96 0.90 1.02AD-20303-b2 0.11 0.07 0.07 0.76 0.64 0.72 AD-20304-b2 0.86 0.85 0.811.03 0.81 1.07 AD-20305-b2 1.03 1.04 1.04 1.08 0.91 1.06 AD-20306-b20.96 0.85 0.89 1.08 0.85 0.99 AD-20307-b2 1.11 1.02 1.00 1.23 0.84 1.07AD-20308-b2 1.03 1.16 1.06 1.15 0.80 1.18 AD-20309-b2 1.06 0.92 1.051.00 0.95 1.16 AD-20310-b2 1.12 0.93 1.12 0.99 0.87 1.17 AD-20311-b21.06 0.81 0.87 1.13 0.83 1.10 AD-20312-b2 0.36 0.33 0.35 0.86 0.83 0.81AD-20313-b2 0.30 0.09 0.23 0.77 1.23 0.69 AD-20314-b2 0.80 0.65 0.781.08 0.83 1.17 AD-20315-b2 0.21 0.14 0.16 0.86 0.79 0.85 AD-20316-b20.57 0.50 0.54 1.09 0.79 1.15 AD-20317-b2 0.89 0.95 0.88 1.27 1.10 1.06AD-20318-b2 0.89 0.93 0.84 1.04 1.10 1.15 AD-20319-b2 0.87 0.63 0.971.13 1.06 1.22 AD-20320-b2 0.85 0.56 0.81 1.05 0.96 1.19 AD-20344-b20.67 0.54 0.50 1.05 1.05 1.03 AD-20345-b2 0.77 0.35 0.76 1.01 1.03 1.07AD-20346-b2 1.13 0.96 0.86 1.17 1.08 1.04 AD-20347-b2 0.78 0.94 0.750.95 0.91 1.07 AD-20348-b2 0.90 0.19 0.74 0.77 0.63 0.80 AD-20349-b20.57 0.47 0.57 0.92 0.73 0.82 AD-20350-b2 0.85 0.72 0.88 1.11 0.72 0.92AD-20351-b2 1.04 0.84 1.12 1.16 1.09 1.08 AD-20352-b2 1.12 0.88 0.930.99 1.13 1.27 AD-20353-b2 0.77 0.53 0.62 1.01 0.81 1.10 AD-20354-b20.78 0.49 0.77 0.99 0.85 1.10 AD-20355-b2 1.13 0.53 0.85 0.97 0.91 1.06AD-20356-b2 0.72 0.63 0.77 0.95 0.81 1.05 AD-20357-b2 1.31 0.98 0.970.96 0.89 1.25 AD-20358-b2 1.09 0.76 1.03 0.92 0.81 1.16 AD-20359-b21.08 1.06 1.17 1.23 0.95 1.03 AD-20360-b2 1.04 1.10 1.04 1.28 1.00 0.90AD-20361-b2 0.93 0.84 0.95 1.03 0.88 1.04 AD-20362-b2 0.40 0.21 0.331.23 0.64 1.05 AD-20363-b2 1.16 0.79 0.98 1.05 0.63 1.25 AD-20364-b20.40 0.21 0.25 0.96 0.42 0.91 AD-20365-b2 0.22 0.12 0.17 0.62 0.57 0.50AD-20366-b2 0.46 0.20 0.31 0.95 0.93 0.84 AD-20367-b2 1.00 0.59 0.831.00 0.71 0.99 AD-20368-b2 1.20 0.50 1.12 1.15 0.87 0.97 AD-20369-b21.02 0.82 0.90 0.88 0.88 0.85 AD-20370-b2 1.04 0.78 0.83 1.03 0.88 0.96AD-20371-b2 1.19 0.71 1.09 1.08 0.83 0.98 AD-20372-b2 1.15 0.73 0.971.07 0.97 0.85 AD-20373-b2 0.20 0.15 0.16 0.74 0.54 0.78 AD-20374-b20.70 0.43 0.58 1.10 0.67 1.04 AD-20375-b2 0.85 0.65 0.72 1.21 0.81 1.06AD-20376-b2 0.13 0.07 0.09 0.48 0.25 0.37 AD-20377-b2 0.35 0.19 0.210.88 0.71 0.78 AD-20378-b2 1.17 0.94 1.19 1.03 0.84 0.98 AD-20379-b20.68 0.43 0.60 0.98 0.77 0.95 AD-20380-b2 0.64 0.43 0.67 1.01 0.81 0.97AD-20381-b2 1.40 1.15 1.49 1.19 0.89 1.13 AD-20382-b2 1.06 0.73 0.971.00 0.94 0.91 AD-20383-b2 1.00 0.95 1.02 1.02 0.94 1.00 AD-20384-b21.05 0.81 0.86 0.99 1.07 0.91 AD-20385-b2 1.15 0.79 0.97 1.19 0.78 1.10AD-20386-b2 0.17 0.04 0.12 0.37 0.32 0.31 AD-20387-b2 0.69 0.38 0.451.06 0.81 0.93 AD-20388-b2 0.66 0.59 0.45 0.98 0.80 0.89 AD-20389-b20.11 0.07 0.08 0.76 0.56 0.67 AD-20390-b2 0.47 0.46 0.41 1.11 0.96 1.12AD-20391-b2 0.07 0.07 0.05 0.53 0.41 0.42 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AD-20692-b2 1.22 1.85 1.31 1.02 1.05 1.46 AD-20693-b21.27 0.32 1.08 1.06 1.01 1.05 AD-20694-b2 0.46 0.12 0.42 0.68 0.45 0.70AD-20695-b2 0.41 0.14 0.36 0.74 0.75 0.73 AD-20696-b2 0.84 1.09 0.821.08 1.06 1.11 AD-20697-b2 1.34 0.67 1.15 1.25 1.02 1.02 AD-20698-b21.25 0.74 0.99 1.11 0.92 1.09 AD-20699-b2 0.97 0.50 1.08 1.16 0.91 1.04AD-20700-b2 0.45 0.21 0.24 1.10 0.81 0.91 AD-20701-b2 0.87 0.30 0.821.14 1.01 1.06 AD-20702-b2 0.36 0.08 0.26 0.53 0.42 0.54 AD-20703-b21.02 0.44 0.92 1.17 0.92 1.01 AD-20704-b2 1.38 0.82 1.14 1.09 1.02 1.11AD-20705-b2 0.78 0.35 0.65 1.05 0.99 1.10 AD-20706-b2 0.70 0.33 0.640.87 0.83 0.80 AD-20707-b2 0.13 0.07 0.14 0.54 0.43 0.55 AD-20708-b20.30 0.12 0.22 1.08 1.08 1.08 AD-20709-b2 0.38 0.19 0.25 0.90 0.92 0.91AD-20710-b2 1.00 0.47 1.07 1.15 1.01 1.12 AD-20711-b2 0.71 0.44 0.681.16 0.99 1.13 AD-20712-b2 1.01 0.71 1.06 0.67 0.47 0.62 AD-20713-b20.61 0.22 0.44 1.17 0.79 0.92 AD-20714-b2 0.49 0.07 0.39 0.56 0.71 0.68AD-20715-b2 0.37 0.08 0.19 1.12 0.94 1.00 AD-20716-b2 0.20 0.07 0.120.87 0.58 0.74 AD-20717-b2 0.81 0.62 0.71 1.18 0.86 0.92 AD-20718-b20.30 0.16 0.19 1.21 0.65 0.75 AD-20719-b2 0.57 0.53 0.58 0.90 0.91 0.91AD-20720-b2 0.57 0.51 0.49 0.85 0.92 0.83 AD-20721-b2 0.67 0.49 0.560.85 0.91 0.86 AD-20722-b2 0.70 0.51 0.66 0.92 0.93 0.89 AD-20723-b20.63 0.48 0.66 0.96 1.03 1.01 AD-20725-b2 0.68 0.66 0.75 0.90 0.99 1.02AD-20726-b2 0.71 0.65 0.64 1.00 0.95 1.07 AD-20727-b2 0.79 0.76 0.760.94 0.99 0.96 AD-20728-b2 0.25 0.26 0.26 0.91 1.03 0.90 AD-20729-b20.72 0.67 0.69 1.15 1.05 0.86 AD-20731-b2 0.58 0.48 0.44 1.11 1.16 1.09AD-20732-b2 0.55 0.51 0.45 0.83 1.33 0.80 AD-20733-b2 0.62 0.71 0.641.07 0.98 1.00 AD-20734-b2 0.79 0.66 0.79 0.95 0.91 1.06 AD-20735-b20.76 0.70 0.69 1.05 1.06 1.04 AD-20736-b2 0.73 0.71 0.77 1.01 0.91 1.07AD-20737-b2 0.98 0.93 1.09 1.17 0.92 1.04 AD-20738-b2 0.94 0.93 0.961.00 1.02 1.09 AD-20739-b2 1.15 0.87 1.19 1.04 0.95 1.00 AD-20740-b20.69 0.65 0.66 1.09 1.00 1.03 AD-20741-b2 1.14 0.87 1.04 1.06 1.08 1.00AD-20742-b2 0.05 0.06 0.06 0.61 0.47 0.52 AD-20743-b2 0.73 1.02 0.740.94 1.09 0.93 AD-20744-b2 0.34 0.27 0.34 1.05 1.08 1.04 AD-20745-b20.41 0.56 0.57 1.00 0.94 1.04 AD-20746-b2 0.69 0.90 0.79 0.89 0.93 0.89AD-20747-b2 0.82 0.81 0.88 1.05 1.01 1.04 AD-20748-b2 0.89 0.89 0.800.87 1.11 1.02 AD-20749-b2 0.44 0.46 0.47 1.03 0.96 0.95 AD-20750-b20.65 0.65 0.68 1.03 0.95 1.02 AD-20751-b2 0.79 0.79 0.82 1.03 0.94 0.98AD-20752-b2 0.42 0.33 0.45 0.86 0.77 0.87 AD-20753-b2 0.95 1.03 1.091.00 1.13 0.97 AD-20755-b2 0.64 0.51 0.54 1.19 1.02 1.13 AD-20756-b20.71 0.61 0.61 1.03 1.10 1.04 AD-20757-b2 0.82 0.78 0.80 1.12 1.15 0.99AD-20758-b2 0.71 0.67 0.72 0.85 0.97 1.05 AD-20759-b2 0.66 0.63 0.670.92 0.94 1.06 AD-20761-b2 0.66 0.65 0.66 0.85 0.87 0.86 AD-20762-b20.78 0.84 0.77 0.89 0.80 0.88 AD-20763-b2 1.06 0.80 1.07 0.91 1.04 0.90AD-20764-b2 0.93 0.88 0.98 0.89 0.83 0.84 AD-20765-b2 0.11 0.07 0.100.54 0.47 0.46 AD-20767-b2 0.45 0.56 0.51 0.82 0.87 0.84 AD-20768-b20.67 0.62 0.66 0.94 1.04 0.95 AD-20769-b2 1.16 1.05 0.99 1.15 1.14 0.98AD-20770-b2 0.64 0.47 0.66 1.01 1.16 1.02 AD-20771-b2 0.92 0.86 0.920.91 0.87 0.90 AD-20772-b2 0.68 0.58 0.66 1.09 1.08 0.97 AD-20774-b20.89 0.86 0.80 1.21 0.95 0.94 AD-20775-b2 0.68 0.54 0.65 1.07 1.01 1.10AD-20776-b2 0.92 0.85 0.91 1.13 1.08 1.03 AD-20777-b2 0.84 0.71 0.881.08 0.92 1.05 AD-20778-b2 0.92 0.90 0.88 1.14 0.96 0.97 AD-20780-b21.06 0.96 1.04 1.02 1.04 1.13 AD-20781-b2 1.06 1.10 1.09 1.11 0.99 1.12AD-20782-b2 0.98 1.01 1.12 1.19 1.14 1.02 AD-20783-b2 1.16 0.93 1.121.10 1.00 1.04 AD-20784-b2 0.38 0.23 0.26 1.08 1.02 0.98 AD-20786-b20.56 0.32 0.43 1.02 0.97 0.91 AD-20787-b2 0.81 0.86 0.72 1.04 0.93 1.00AD-20788-b2 1.07 0.84 1.09 0.94 1.02 1.03 AD-20789-b2 1.01 0.73 1.061.07 0.97 1.06 AD-20790-b2 0.85 0.61 0.89 0.82 0.99 0.91 AD-20792-b21.03 0.99 1.04 1.07 1.09 1.05 AD-20793-b2 1.24 0.93 1.02 1.00 1.05 0.99AD-20794-b2 1.05 0.94 1.20 1.02 1.02 1.00 AD-20795-b2 1.01 0.93 1.081.01 1.04 0.98 AD-20796-b2 1.25 1.04 1.08 0.96 1.09 0.94 AD-20797-b21.01 1.01 0.83 1.03 1.09 1.00 AD-20798-b2 0.91 0.96 1.00 1.01 1.11 0.96AD-20799-b2 0.87 0.77 0.85 1.01 1.21 1.02 AD-20800-b2 1.01 0.99 1.041.07 1.08 0.90 AD-20801-b2 0.90 0.80 0.88 0.98 0.78 0.80 AD-20868-b20.88 0.71 0.85 1.20 0.82 0.81 AD-20869-b2 0.59 0.50 0.52 1.16 1.02 0.92AD-20870-b2 0.66 0.62 0.63 1.29 0.80 0.88 AD-20871-b2 1.03 0.97 1.041.16 0.90 0.97 AD-20872-b2 0.86 0.66 0.80 1.23 0.99 0.93 AD-20873-b20.91 0.98 0.90 1.23 0.85 0.86

Example 7 In Vivo and PBMC Analysis of RNAi Agents

Selected RNAi agents to HSF1 are analyzed in a PBMC (peripheral bloodmononuclear cell) assay to estimate immunogenicity; and in vivo in Hep3B(primary liver xenograft) subcutaneous tumors in nude mice. RNAi agentsare delivered in vivo in a lipid nanoparticle. Single dosages of 5 mg/kgor 3 mg/kg were delivered, and animals sacrificed and tissues collected72 hours after dosage. The results are shown below in Table 8.

The Duplex ID indicates the various names for the RNAi agent;AD-XXXXX/AD-XXXXX=dTdT/uu modified version of the same siRNA coresequence indicated. (XXXXX) indicates the duplex ID for the uu sequence.The Target Seq (sequence) and position in the human and cyno(cynomolgus) HSF1 genes are given; “na” indicates that the givensequence is not in the cynomolgus gene. PBMC Result (IFN-α, TNF-α)indicates whether or not a given RNAi elicited an immune response, asindicated by a measured induction of TNF-α or IFN-α. “quiet” indicatesthat a given RNAi agent did not illicit an immune response. In thecolumns labeled “In vivo Hep3B,” the numbers indicate the per centknockdown of HSF1 expression.

TABLE 8 In vivo and PBMC analysis of RNAi agents PBMC In Result vivo Invivo (IFN- Hep- Hep- SEQ α, 3B 3B Duplex ID Pos Pos TNF- 5 mg/kg, 3mg/kg, ID Target Seq (=SS 19-mer) NO: Human Cyno α) 72 h 72 h AD-UUGAGAACAUCAAGAGGAA 3195 505 na Quiet 50% 52% 20403/ & AD-30070 70%(505) AD- CCCUGAAGAGUGAAGACAU 3196 541 532 Quiet 50% 40% 20437/ AD-36969AD-20438 CCUGAAGAGUGAAGACAUA 3197 542 533 Quiet 50% 62% AD-20439CUGAAGAGUGAAGACAUAA 3198 543 534 Quiet 47% 46% AD- UGAAGAGUGAAGACAUAAA3199 544 535 Quiet 60-70% 70% 20487/ AD-30071 (544) AD-AAGAGUGAAGACAUAAAGA 3200 546 537 Mild 39% 20489/ TNF-α AD-36970induction AD-20490 AGAGUGAAGACAUAAAGAU 3201 547 538 Quiet 42% AD-20491GAGUGAAGACAUAAAGAUC 3202 548 539 Quiet 40% AD-20548 GCAGAAGCAUGCCCAGCAA3203 698 689 Quiet 40% AD- AGCUCAUUCAGUUCCUGAU 3204 733 724 Quiet 60%50% 20560/ AD-37739 (733) AD-20562 CUCAUUCAGUUCCUGAUCU 3205 735 726Quiet 50% 50% (735) AD- UCAUUCAGUUCCUGAUCUC 3206 736 727 Quiet 34%20563/ AD-36973 AD- CAUUCAGUUCCUGAUCUCA 3207 737 728 Quiet 57% 65%20564/ AD-36971 AD-20578 UCUCACUGGUGCAGUCAAA 3208 751 742 Quiet 32%AD-20626 UGGUGCAGUCAAACCGGAU 3209 757 748 Quiet 36% AD-20627GGUGCAGUCAAACCGGAUC 3210 758 749 Quiet 50% 40% (758) AD-20644GAACGACAGUGGCUCAGCA 3211 809 800 Quiet 17% AD-20648 GACAGUGGCUCAGCACAUU3212 813 804 Quiet 27% AD- GUGGCUCAGCACAUUCCAU 3213 817 808 Quiet 28%20652/ AD-36972 AD-20660 GCACAUUCCAUGCCCAAGU 3214 825 816 Quiet 50% 40%(825) AD-20694 GCGGGAGCAUAGACGAGAG 3215 1012 1003 Quiet 24% AD-20707ACUUGGAUGCUAUGGACUC 3216 1330 1321 Quiet 50% (1330) AD-20730UGCUGAGCAGCCACGGCUU 3217 1372 1363 TNF-α 50% 47% (1372) induction

Thus, as shown in the table above, several RNAi agents do not elicit animmune response in the PBMC assay (indicated by “quiet”). Several RNAiagents also elicited at least a 40% HSF1 expression knock-down.AD-20403/30070, AD-20437/AD-36969, AD-20438, AD-20439, AD-20487/AD-30071(544), AD-20548, AD-20560/AD-37739 (733), AD-20562 (735),AD-20564/AD-36971, AD-20627 (758), AD-20660 (825), AD-20707 (1330), andAD-20730 (1372) are chosen for lead optimization.

Example 8 5′-End Modifications

This example describes screening multiple chemistries to identify siRNAsthat retain or improve activity relative to parent and that arepotentially more stable.

A two-phase approach is used. The first phase involves identifying anoptimal 5′ end cap for S (sense) strand inactivation. End-caps used are:Inverted dT, L-sugar, and C-6-alkyl. These are illustrated in FIG. 1.Each cap is tested for its ability to block siRNA activity when on the5′-end of antisense (AS) but maintain activity when on the 5′-end of thesense strand using 4 potent HSF1 siRNAs for evaluation. IC50 (EC50) inHeLa cells is determined.

In the second phase, multiple chemical motifs are tested on a larger setof HSF1 siRNAs in context of the best 5′-end cap found in the firstphase.

S (sense) modifications include the following:

-   Endolight, wherein all pyrmidines are modified to 2′-OMe with 2′-OMe    uu overhang, and-   All ribonucleotides are used with 2′-OMe uu overhang.

AS modifications include:

-   All pyrimidines are modified with to 2′-OMe, with a 2′-OMe uu    overhang,-   Endolight+2′-OMe on 5 3′-terminal bases with 2′-OMe uu overhang-   Endolight+2′-OMe at only pyrmidines of 5 3′ terminal bases, with    2′-OMe uu overhang.

Table 9A and Table 9B provide examples of 5′-end modificationscontemplated within the scope of the invention. As depicted in thetables, IC50 I and IC50 II indicate the results of two determinations ofIC50 (EC50). The results indicate nM concentrations.

TABLE 9A Antisense modifications SEQ ID Duplex ID Sequence NO: IC50 IIC50 II dTdT/uu AD-20437.4 cccuGAAGAGuGAAGAcAudTdT 3218 0.014 0.016AUGUCUUcACUCUUcAGGGdTdT 3219 AD-20487.7 uGAAGAGuGAAGAcAuAAAdTdT 32200.003 0.004 UUuAUGUCUUcACUCUUcAdTdT 3221 AD-20489.2AAGAGuGAAGAcAuAAAGAdTdT 3222 0.005 0.005 UCUUuAUGUCUUcACUCUUdTdT 3223AD-20560.4 AGcucAuucAGuuccuGAudTdT 3224 0.013 0.012AUcAGGAACUGAAUGAGCUdTdT 3225 Inverted AD-37718.1 cccuGAAGAGuGAAGAcAuuu3226 0.126 0.220 dT (idT)AUGUCUUcACUCUUcAGGGuu 3227 AD-37721.1uGAAGAGuGAAGAcAuAAAuu 3228 1.800 >10 (idT)UUuAUGUCUUcACUCUUcAuu 3229AD-37724.1 AAGAGuGAAGAcAuAAAGAuu 3230 0.473 0.737(idT)UCUUuAUGUCUUcACUCUUuu 3231 AD-37727.1 AGcucAuucAGuuccuGAuuu3232 >10 3.202 (idT)AUcAGGAACUGAAUGAGCUuu 3233 L-sugar AD-37730.1cccuGAAGAGuGAAGAcAuuu 3234 0.240 0.326 AbUGUCUUcACUCUUcAGGGuu 3235AD-37733.1 uGAAGAGuGAAGAcAuAAAuu 3236 >10 5.649 UbUuAUGUCUUcACUCUUcAuu3237 AD-37736.1 AAGAGuGAAGAcAuAAAGAuu 3238 0.157 0.254UbCUUuAUGUCUUcACUCUUuu 3239 AD-37740.1 AGcucAuucAGuuccuGAuuu 3240 0.0280.064 AbUcAGGAACUGAAUGAGCUuu 3241 C-6-alkyl AD-37719.1cccuGAAGAGuGAAGAcAuuu 3242 0.471 0.451 Q128AUGUCUUcACUCUUcAGGGuu 3243AD-37722.1 uGAAGAGuGAAGAcAuAAAuu 3244 0.087 0.031Q128UUuAUGUCUUcACUCUUcAuu 3245 AD-37725.1 AAGAGuGAAGAcAuAAAGAuu 32460.013 0.017 Q128UCUUuAUGUCUUcACUCUUuu 3247 AD-37728.1AGcucAuucAGuuccuGAuuu 3248 0.006 0.005 Q128AUcAGGAACUGAAUGAGCUuu 3249

TABLE 9B Sense modifications SEQ ID Duplex ID Sequence NO: IC50 I IC50II dTdT/uu AD-36969.2 cccuGAAGAGuGAAGAcAuuu 3250 0.014 0.006AUGUCUUcACUCUUcAGGGuu 3251 AD-30071.2 uGAAGAGuGAAGAcAuAAAuu 3252 0.0030.006 UUuAUGUCUUcACUCUUcAuu 3253 AD-36970.2 AAGAGuGAAGAcAuAAAGAuu 32540.015 0.011 UCUUuAUGUCUUcACUCUUuu 3255 AD-37739.1 AGcucAuucAGuuccuGAuuu3256 0.804 0.644 AUcAGGAACUGAAUGAGCUuu 3257 Inverted AD-37731.1(idT)cccuGAAGAGuGAAGAcAuuu 3258 0.012 0.008 dT AUGUCUUcACUCUUcAGGGuu3259 AD-37734.1 (idT)uGAAGAGuGAAGAcAuAAAuu 3260 0.009 0.009UUuAUGUCUUcACUCUUcAuu 3261 AD-37737.1 (idT)AAGAGuGAAGAcAuAAAGAuu 32620.003 0.005 UCUUuAUGUCUUcACUCUUuu 3263 AD-37741.1(idT)AGcucAuucAGuuccuGAuuu 3264 0.005 0.008 AUcAGGAACUGAAUGAGCUuu 3265L-sugar AD-37720.1 CbccuGAAGAGuGAAGAcAuuu 3266 0.004 0.004AUGUCUUcACUCUUcAGGGuu 3267 AD-37723.1 UbGAAGAGuGAAGAcAuAAAuu 3268 0.0100.010 UUuAUGUCUUcACUCUUcAuu 3269 AD-37726.1 AbAGAGuGAAGAcAuAAAGAuu 32700.003 0.003 UCUUuAUGUCUUcACUCUUuu 3271 AD-37729.1 AbGcucAuucAGuuccuGAuuu3272 0.006 0.010 AUcAGGAACUGAAUGAGCUuu 3273 C-6-alkyl AD-37732.1Q128CccuGAAGAGuGAAGAcAuuu 3274 0.014 0.005 AUGUCUUcACUCUUcAGGGuu 3275AD-37735.1 Q128UGAAGAGuGAAGAcAuAAAuu 3276 0.144 UUuAUGUCUUcACUCUUcAuu3277 AD-37738.1 Q128AAGAGuGAAGAcAuAAAGAuu 3278 0.008 0.087UCUUuAUGUCUUcACUCUUuu 3279 AD-37742.1 Q128AGcucAuucAGuuccuGAuuu 32800.010 0.020 AUcAGGAACUGAAUGAGCUuu 3281

Note that in the above tables, a suffix such as 0.1, 0.2, 0.3, 0.4, etc.indicates a variant of a given RNAi agent. Thus, AD-20437.4 andAD-20437, for example, have the same sequence, though they may vary inmodifications, caps, etc. Thus, any references to (includingdescriptions of various embodiments related to) AD-20437.4; AD-20487.7;AD-20489.2; AD-20560.4; AD-37718.1; AD-37721.1; AD-37724.1; AD-37727.1;AD-37730.1; AD-37733.1; AD-37736.1; AD-37740.1; AD-37719.1; AD-37722.1;AD-37725.1; AD-37728.1; AD-30071.2; AD-36969.2; AD-36970.2; AD-37718.1;AD-37719.1; AD-37720.1; AD-37721.1; AD-37722.1; AD-37723.1; AD-37724.1;AD-37725.1; AD-37726.1; AD-37727.1; AD-37728.1; AD-37729.1; AD-37730.1;AD-37731.1; AD-37732.1; AD-37733.1; AD-37734.1; AD-37735.1; AD-37736.1;AD-37737.1; AD-37738.1; AD-37739.1; AD-37740.1; AD-37741.1; AD-37742.1;also refer, respectively, to: AD-20437; AD-20487; AD-20489.2; AD-20560;AD-37718; AD-37721; AD-37724; AD-37727; AD-37730; AD-37733; AD-37736;AD-37740; AD-37719; AD-37722; AD-37725; AD-37728; AD-30071; AD-36969;AD-36970; AD-37718; AD-37719; AD-37720; AD-37721; AD-37722; AD-37723;AD-37724; AD-37725; AD-37726; AD-37727; AD-37728; AD-37729; AD-37730;AD-37731; AD-37732; AD-37733; AD-37734; AD-37735; AD-37736; AD-37737;AD-37738; AD-37739; AD-37740; AD-37741; and AD-37742. Thus, any groupingof siRNAs of overlapping sequences comprising AD-20437 also comprisesAD-20437.4. This is true of other groupings comprising any variantsequence; thus, any grouping of overlapping siRNAs of a given sequencealso comprises siRNAs comprising a variant of that sequence, e.g., withmodifications and/or caps.

The inverted dT (idT) and L-sugar are found to reduce activityapproximately 10- to 100-fold when the antisense strand is modified;there is minimal impact when these modifications are placed on the5′-end of the sense strand.

Unless defined otherwise, the technical and scientific terms used hereinhave the same meaning as that usually understood by a specialistfamiliar with the field to which the disclosure belongs.

Unless indicated otherwise, all methods, steps, techniques andmanipulations that are not specifically described in detail can beperformed and have been performed in a manner known per se, as will beclear to the skilled person. Reference is for example again made to thestandard handbooks and the general background art mentioned herein andto the further references cited therein. Unless indicated otherwise,each of the references cited herein is incorporated in its entirety byreference.

Claims to the invention are non-limiting and are provided below.

Although particular embodiments and claims have been disclosed herein indetail, this has been done by way of example for purposes ofillustration only, and is not intended to be limiting with respect tothe scope of the appended claims, or the scope of subject matter ofclaims of any corresponding future application. In particular, it iscontemplated by the inventors that various substitutions, alterations,and modifications may be made to the disclosure without departing fromthe spirit and scope of the disclosure as defined by the claims. Thechoice of nucleic acid starting material, clone of interest, or librarytype is believed to be a matter of routine for a person of ordinaryskill in the art with knowledge of the embodiments described herein.Other aspects, advantages, and modifications considered to be within thescope of the following claims. Those skilled in the art will recognizeor be able to ascertain, using no more than routine experimentation,many equivalents of the specific embodiments of the invention describedherein. Such equivalents are intended to be encompassed by the followingclaims. Redrafting of claim scope in later filed correspondingapplications may be due to limitations by the patent laws of variouscountries and should not be interpreted as giving up subject matter ofthe claims.

1. A composition comprising an RNAi agent to HSF1 comprising a firststrand and a second strand, wherein the sequence of the first strandcomprises at least 15 contiguous nucleotides differing by 0, 1, 2, or 3nucleotides from the sequence of SEQ ID NO: 679, 680, 681, 682 or 683,and wherein the first and/or second strand are modified or unmodified.2. The composition of claim 1, wherein the composition further comprisesa second RNAi agent to HSF1.
 3. The composition of claim 1, wherein thefirst strand and the second strand are both about 19 to about 23nucleotides in length.
 4. The composition of claim 1, wherein the RNAiagent comprises a modified sugar backbone, a phosphorothioate linkage,or a 2′-modified nucleotide.
 5. The composition of claim 1, wherein theRNAi agent comprises a 2′-modification selected from the groupconsisting of: 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl,2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP),2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl(2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), and2′-O—N-methylacetamido (2′-O-NMA).
 6. The composition of claim 1,wherein the RNAi agent comprises a blunt end or an overhang having 1 to4 unpaired nucleotides.
 7. The composition of claim 1, wherein the RNAiagent is ligated to one or more diagnostic compound, reporter group,cross-linking agent, nuclease-resistance conferring moiety, natural orunusual nucleobase, lipophilic molecule, cholesterol, lipid, lectin,steroid, uvaol, hecigenin, diosgenin, terpene, triterpene,sarsasapogenin, Friedelin, epifriedelanol-derivatized lithocholic acid,vitamin, carbohydrate, dextran, pullulan, chitin, chitosan, syntheticcarbohydrate, oligo lactate 15-mer, natural polymer, low- ormedium-molecular weight polymer, inulin, cyclodextrin, hyaluronic acid,protein, protein-binding agent, integrin-targeting molecule,polycationic, peptide, polyamine, peptide mimic, and/or transferrin. 8.A composition comprising an RNAi agent to HSF1 comprising a first strandand a second strand, wherein the sequence of the first strand comprisesthe sequence of SEQ ID NO: 679, 680, 681, 682 or 683 and/or the sequenceof the second strand comprises the sequence of SEQ ID NO: 167, 168, 169,170 or 171, and wherein the first and/or second strand are modified orunmodified.
 9. The composition of claim 8, wherein the compositionfurther comprises a second RNAi agent to HSF1.
 10. The composition ofclaim 8, wherein the first strand and the second strand are both 19 to23 nucleotides in length.
 11. The composition of claim 8, wherein theRNAi agent comprises a modified sugar backbone, a phosphorothioatelinkage, or a 2′-modified nucleotide.
 12. The composition of claim 8,wherein the RNAi agent comprises a 2′-modification selected from thegroup consisting of: 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl,2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP),2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl(2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), and2′-O—N-methylacetamido (2′-O-NMA).
 13. The composition of claim 8,wherein the RNAi agent comprises a blunt end or an overhang having 1 to4 unpaired nucleotides.
 14. The composition of claim 8, wherein the RNAiagent is ligated to one or more diagnostic compound, reporter group,cross-linking agent, nuclease-resistance conferring moiety, natural orunusual nucleobase, lipophilic molecule, cholesterol, lipid, lectin,steroid, uvaol, hecigenin, diosgenin, terpene, triterpene,sarsasapogenin, Friedelin, epifriedelanol-derivatized lithocholic acid,vitamin, carbohydrate, dextran, pullulan, chitin, chitosan, syntheticcarbohydrate, oligo lactate 15-mer, natural polymer, low- ormedium-molecular weight polymer, inulin, cyclodextrin, hyaluronic acid,protein, protein-binding agent, integrin-targeting molecule,polycationic, peptide, polyamine, peptide mimic, and/or transferrin. 15.A method of inhibiting the expression of HSF1 in an individual,comprising the step of administering to the individual a compositioncomprising a pharmaceutically acceptable carrier and a therapeuticallyeffective amount of an RNAi agent to HSF1 comprising a first strand anda second strand, wherein the sequence of the first strand comprises thesequence of SEQ ID NO: 679, 680, 681, 682 or 683, and wherein the firstand/or second strand are modified or unmodified, the method optionallyfurther comprising the step of administering a second RNAi agent toHSF1.
 16. The method of claim 15, wherein the individual is afflictedwith an HSF1-related disease.
 17. The method of claim 16, wherein theHSF1-related disease is cancer or a viral disease.
 18. The method ofclaim 17, wherein the HSF1-related disease is cancer selected from thelist of bladder, bone, breast, cervical, colon, colorectal, endometrial,fibrosarcoma, gastric, haematopoietic, intestine, kidney, liver, lung,lymphoma, neuroectodermal, neuroblastoma, Ewing's sarcoma, osteosarcoma,ovary, pancreas, pleura, prostate, skin, squamous cell, stomach, andtesticular cancers, leukemia, promyelocytic leukemia, and Hodgkin'sdisease.
 19. The method of claim 17, wherein the method furthercomprises the step of administering an additional cancer or viraldisease treatment.
 20. The method of claim 17, where the HSF1-relateddisease is a viral disease selected from the list of viral diseasesmediated in whole or in part by adenovirus, herpes simplex virus, humancytomegalovirus, HTLV-1, SV40, polyoma virus, HIV, and/or Epstein-Barrvirus.
 21. The method of claim 16, wherein the HSF1-related disease isan autoimmune disease.
 22. The method of claim 16, wherein theHSF1-related disease is lupus or rheumatoid arthritis.